Study Guide. National Registry of Microbiologists. The National Registry of Microbiologists 2004 American Society for Microbiology

Transcription

1 Study Guide National Registry of Microbiologists The National Registry of Microbiologists 2004 American Society for Microbiology

2 TABLE OF CONTENTS Examination Content...3 Clinical and Public Health Microbiology RM(NRM) Task List...4 Sample Questions...5 Resources...9 Consumer Products and Quality Assurance Microbiology RM(NRM) Task List...10 Sample Questions...12 Resources...17 Public Health and Medical Laboratory Microbiology SM(NRM) Task List...19 Sample Questions...20 Resources...25 Consumer and Industrial Microbiology SM(NRM) Task List...26 Sample Questions...28 Resources...32 Biological Safety Microbiology SM(NRM) Task List...34 Sample Questions...36 Resources

3 EXAMINATION FORMAT AND RESOURCES Question Format and Content The examination consists of 150 multiple-choice questions. Each question has only one correct answer. Questions have a stem and up to five possible responses. Questions are updated and re-evaluated annually. Questions are classified by domain and then by task. The examination will have at least one question from each task. A task list for each specialty examination is included in the study guide. The number of questions from each domain is listed next to the domain s description on the task list for each examination. Scoring The NRM uses a criterion-referencing system for determining examination scores. This method sets a standard of performance in absolute, not relative, terms. As a result, you are not graded on a curve and do not compete against other examinees. If you perform at or above the established criterion standard, you pass the examination and are certified. The number of correct answers determines your score. One point is given for each correct answer. There is no penalty for guessing; therefore, it is in your best interest to answer all questions. Answer sheets are scored electronically. About the Resource Lists The resources listed at the end of each Task List are NOT meant to be comprehensive guides to the examinations. They are merely suggested references for review. If you have questions about any of the material, please do not hesitate to contact the NRM office. 3

4 Clinical and Public Health (C/PH) Task List I. LABORATORY INSTRUMENTS AND EQUIPMENT (20 questions) 1. Use and understand the principles of a steam autoclave. 2. Use and understand the theory of ambient air, carbon dioxide, and anaerobic incubators. 3. Use and understand the principles of laboratory equipment such as pipettes, balances, and ph meters in preparation of media and reagents. 4. Use a biosafety and laminar flow cabinet and fume hood. 5. Use light-field, dark-field, and fluorescence microscopes. 6. Use and understand systems designed to detect, identify, and quantify microorganisms (e.g., BACTEC, Vitek, API). 7. Use and understand systems designed for antimicrobial susceptibility testing (e.g., Microscan, Vitek, E-test). 8. Select and understand principles of stains and media used in identification procedures for bacteria, fungi, parasites, and viruses. II. RAPID SCREENING AND IDENTIFICATION (18 questions) 9. Perform serological tests for syphilis. 10. Select, perform, and interpret techniques for the rapid identification of bacteria based on standard biochemical and enzymatic reactions (e.g., spot tests). 11. Select, perform, and interpret techniques for the identification of viruses. 12. Interpret results of immunological procedures or tests such as agglutination, enzyme immunoassay (EIA), enzyme detection, and fluorescent antibody (FA). 13. Understand theory and application of polymerase chain reaction (PCR) and DNA probes. 4 III. SAMPLE COLLECTION AND TRANSPORT (15 questions) 14. Select appropriate methods for sample collection, transport, and storage. 15. Select and evaluate the acceptability of specimens. 16. Select appropriate test procedures and media for the type of specimen received. 17. Select appropriate incubation conditions (e.g., time, temperature, atmosphere) for the type of specimen received. IV. LABORATORY PROCEDURES (63 questions) 18. Isolate and identify aerobic, gram-positive bacilli. 19. Isolate and identify Bacteroides and other gram-negative, anaerobic bacilli. 20. Isolate and identify Clostridium, Propionibacterium, or other gram-positive, anaerobic bacteria. 21. Isolate and identify Chlamydia, Mycoplasma, and Ureaplasma. 22. Isolate and identify Salmonella, Shigella, Yersinia, and Escherichia coli from enteric specimens. 23. Isolate and identify members of the Enterobacteriaceae family. 24. Isolate and identify fastidious bacteria such as Pasteurella, Francisella, Brucella, Legionella, and Bordetella. 25. Isolate and identify Haemophilus. 26. Isolate and identify Mycobacterium. 27. Isolate and identify Neisseria and Moraxella. 28. Isolate and identify Nocardia, Streptomyces, and Actinomyces. 29. Isolate and identify Pseudomonas. 30. Isolate and identify Staphylococcus. 31. Isolate and identify Streptococcus and Enterococcus. 32. Isolate and identify Vibrio and Aeromonas. 33. Isolate and identify Campylobacter. 34. Isolate and identify the agents of superficial, cutaneous, subcutaneous, and dermatophyte mycoses. 35. Detect and identify dimorphic fungi. 36. Detect and identify opportunistic fungi (e.g., yeasts and molds). 37. Detect and identify intestinal protozoa. 38. Detect and identify urogenital protozoa.

5 39. Detect and identify blood and tissue protozoa. 40. Detect and identify intestinal and tissue helminths. 41. Perform broth and agar dilution susceptibility tests. 42. Perform disk diffusion susceptibility tests. 43. Correlate arthropod vectors and modes of transmission with infectious agents. 44. Know basic cell culture techniques for virus isolation and identification. V. LABORATORY OPERATIONS (17 questions) 45. Use appropriate safety equipment and devices (e.g., goggles, gloves, sharps containers). 46. Use appropriate work practices to reduce the risk of acquiring laboratory infections. 47. Dispose of hazardous substances in compliance with Occupational Safety and Health Administration (OSHA) regulations. 48. Know procedures to monitor quality of media, reagents, equipment, and test methodologies. 49. Monitor and evaluate test results and data for accuracy and recognize aberrant results by using manual or computer database systems. 50. Evaluate, report, and correlate results with clinical information. VI. PUBLIC HEALTH (17 questions) 51. Know the modes of transmission and the symptoms of diseases caused by organisms that pose a threat to public health. 52. Know the general procedures for notifying public health authorities of reportable diseases. 53. Know how to contain and dispose of infectious material in the community. C/PH Sample Questions 1. A common indicator used to ensure the anaerobic atmosphere of the GasPak jar is: a. methyl red. b. pheonlphthalein. c. resazurin. d. methylene blue. e. neutral red. Corresponds to task #2. 2. A bright-field microscope s effectiveness in distinguishing two separate objects is defined as its: a. resolving power. b. enlarging power. c. aberration level. d. monochromicity. e. multichromicity. Corresponds to task #5. 3. In the direct immunofluorescence identification of a specific infectious agent, fluorescein is conjugated to: a. goat antihuman globulin. b. antibody specific to the infectious agent. c. rabbit antihuman globulin. d. a carrier protein. e. horseradish peroxidase. Corresponds to task #5. 5

6 4. Most automated identification systems for bacteria and yeasts: a. will not identify to species. b. use conventional reactions and compare them to a database for probability of genus and species. c. use an antigen-antibody reaction specific for a certain organism. d. cannot be used without a computer system. Corresponds to task #6. 5. Which of the following statements is true regarding automated systems that can determine the organism present by use of turbidometric or colorimetric measurement? a. Increased number of organisms present will increase the turbidity of the suspension. b. Increased number of organisms present will decrease the turbidity of the suspension. c. Growth inhibitors in some wells will increase the turbidity of the suspension. d. Growth inhibitors in some wells will increase colorimetric changes. e. Identification of single organism from a mixed culture may be made. Corresponds to task #6. 6. The most important factor in utilizing automated equipment for the identification of microorganisms is: a. quality control using known organisms. b. proper isolation of the organism. c. technique in inoculation of equipment panels. d. interpretation of results. Corresponds to task #6. 7. Which of the following systems used to identify microorganisms are mechanized or automated? a. Enteric-Tek, N/F system b. Minitek, Rapid E c. BACTEC, Bac-T-Screen d. API 20E, Enterotube II e. Micro-ID system, Oxi-Ferm Corresponds to task #6. 8. Auramine and rhodamine can be used in staining: a. Mycoplasma. b. Chlamydia. c. Microsporum. d. Cryptosporidium. e. Actinomyces. Corresponds to task #8. 9. The test which is most often used as a confirmatory test for exposure to the human immunodeficiency virus is: a. Western blot analysis. b. radial immunodiffusion. c. enzyme-linked immunosorbent assay (ELISA). d. direct immunofluorescence. e. counterimmunoelectrophoresis. Corresponds to task # The indirect fluorescent antibody test for immunoglobulin M (IgM) antibody may give false-positive reactions in the presence of: a. IgE. b. complement. c. rheumatoid factor. d. elevated protein. e. IgA. Corresponds to task #12. 6

7 11. Cultures of aerobic bacteria should be shipped: a. in sealed containers. b. in cotton-plugged tubes. c. lyophilized. d. in broth cultures. e. by ground transportation only. Corresponds to task # When clinical microbiologists are asked to identify arthropods or arthropod larvae, ectoparasite specimens should be preserved and shipped for identification in: a. 5% phenol. b. polyvinyl alcohol (PVA). c. 70% ethyl alcohol. d. acetone. e. Benedict s solution. Corresponds to task # Which of the following procedures is correct for the isolation of Bordetella pertussis? a. Nasopharyngeal swabs plated on Regan-Lowe medium b. Transtracheal aspirate plated on Feeley-Gorman agar c. Nasopharyngeal swabs plated on sheep blood agar d. Transtracheal aspirate plated on Middlebrook 7H10 e. Cough plate on chocolate agar Corresponds to task # On Christensen urea agar medium, the rate of urea hydrolysis differentiates: a. Klebsiella from Citrobacter. b. Proteus species from other urease-positive organisms. c. Enterobacter from Citrobacter. d. Morganella morganii from Proteus vulgaris. e. Klebsiella from Enterobacter. Corresponds to task # Francisella tularensis can most rapidly be detected by: a. direct light microscopic examination. b. FA techniques. c. microscopic examination. d. culture on selective media. e. inoculation of laboratory animals. Corresponds to task # On charcoal yeast extract agar, Legionella pneumophila appears: a. light blue with a cut-glass texture. b. white with a brown halo. c. green with a spreading edge. d. pink with a rhizoid edge. e. tan with an entire edge. Corresponds to task # Which of the following is NOT part of the recommended procedure for the isolation of Brucella species? a. Collection of multiple blood specimens b. Inoculation of specimens into a biphasic bottle c. Incubation under anaerobic conditions d. Incubation at 35 C e. Examination of cultures for 20 days Corresponds to task # A very common yeast which may be isolated from urine and has a size of 2 to 3 µm is: a. Trichosporon cutaneum. b. Saccharomyces cerevisiae. c. Rhodotorula rubra. d. Candida (Torulopsis) glabrata. e. Cryptococcus neoformans. Corresponds to task #36. 7

8 19. If incubation of a yeast in serum for two hours at 37 C does not induce germ tubes, the yeast is probably not: a. Cryptococcus neoformans. b. Candida tropicalis. c. Candida glabrata. d. Saccharomyces cerevisiae. e. Candida albicans. Corresponds to task # Which of the following Aspergillus species is most commonly isolated from respiratory specimens? a. flavus b. fumigatus c. terreus d. niger e. nidulans Corresponds to task # Toxoplasma gondii is: a. an obligate intracytoplasmic parasite. b. very host specific. c. gram positive. d. limited to the felines as intermediate hosts. e. found in the blood but cannot cross the placenta. Corresponds to task # A tube dilution to test susceptibility of Haemophilus influenzae to ampicillin is performed. Tube #1 = 1.6 µg of ampicillin/ml Tube #2 = 0.8 µg of ampicillin/ml Tube #3 = 0.4 µg of ampicillin/ml Tube #4 = 0.2 µg of ampicillin/ml Tube #5 = 0.1 µg of ampicillin/ml Tube #6 = no ampicillin Growth is visualized in tubes #3 through 6. The proper report for this result is: a. MIC = 0.4. b. MIC = 0.8. c. minimal bactericidal concentration (MBC) = 0.4. d. MBC = 0.8. e. MBC = 1.6. Corresponds to task # Antibiotics are routinely added to cell culture media to: a. help the cells adhere to the flask. b. protect the cells from bacterial contamination. c. maintain the proper ph. d. help in the identification of infecting viruses. e. interfere with the culturing of clinical specimens. Corresponds to task # Which of the following types of agents that control microflora is the MOST lethal? a. Disinfectant b. Sanitizer c. Antiseptic d. Bactericide Corresponds to task #46. 8

9 25. A biological safety cabinet is LEAST necessary when working with: a. Coccidioides immitis. b. Histoplasma capsulatum. c. Streptobacillus moniliformis. d. Mycobacterium tuberculosis. e. Blastomyces dermatitidis. Corresponds to task # Generation of aerosols can be prevented by: a. centrifuging with covered buckets. b. preparing a slide for gram staining. c. flaming a bacteriologic loop. d. pipetting. e. inoculating laboratory animals. Corresponds to task #47. ANSWERS 1. d 6. b 11. a 16. a 21. a 26. a 2. a 7. c 12. c 17. c 22. b 3. b 8. d 13. a 18. d 23. b 4. b 9. a 14. b 19. e 24. d 5. a 10. c 15. b 20. b 25. b RESOURCES Critical References Books Murray, P.R., E.J. Baron, M.A. Pfaller, F.C. Tenover, and R.H. Yolken (ed.) Manual of clinical microbiology, 8 th ed. ASM Press, Washington, D.C. Baron, E.J., L.R. Peterson, and S.M. Finegold (ed.) Bailey and Scott s diagnostic microbiology, 9 th ed. C.V. Mosby, St. Louis, Mo. Journal Clinical Microbiology Reviews. American Society for Microbiology, Washington, D.C. Helpful References Ciulla, A., and G. Buescher Prentice Hall Health s question and answer review of medical technology/clinical laboratory science, 3 rd ed. Prentice-Hall. 9

10 Consumer Products and Quality Assurance (CP/QA) Task List Specialty examinations consist of 100 general questions and 50 specialty questions. General Tasks I. LABORATORY INSTRUMENTS AND EQUIPMENT (21 questions) 1. Validate, use, and monitor a steam autoclave. 2. Use a ph or conductivity meter. 3. Use various types of microscopes (e.g., light field, dark field, phase contrast, fluorescence). 4. Use filtration equipment for sterilization of solutions. 5. Use spectrophotometric and colorimetric equipment. 6. Use fermentors and/or continuous culture apparatuses. 7. Use laminar flow hood and biosafety cabinets. 8. Use incubation chambers and appropriate controls. II. LABORATORY PREPARATIONS (12 questions) 9. Use general stains (e.g., Gram, nigrosin, spore, Ziehl-Neelsen, Kinyoun, flourescent). 10. Prepare and perform appropriate quality control checks on media from commercial dehydrated materials and supplements. 11. Use general, selective, or differential media for bacteria. 12. Use general, selective, or differential media for fungi. 13. Prepare solutions of known molarity, molality, and normality. 14. Prepare and use buffers. III. LABORATORY PROCEDURES (43 questions) 15. Isolate and identify Bacillus or other gram-positive, aerobic bacilli Isolate and identify Pseudomonas and other oxidative, gramnegative bacilli. 17. Isolate and identify Enterobacteriaceae. 18. Isolate and identify Staphylococcus. 19. Isolate and identify Streptococcus. 20. Perform precipitation, agglutination, immunoassay, and immunoflourescence tests. 21. Perform broth or agar susceptibility tests of antimicrobials. 22. Detect and measure the growth of microorganisms (e.g., by substrate utilization, turbidity, impedance, and rapid methodologies). 23. Determine inactivation rates of microorganisms by chemical and physical means. 24. Use viable plate count procedures. 25. Use most-probable-number technique. 26. Perform tests for water suitability in production systems. 27. Perform phenol coefficient tests or similar tests on disinfectants. 28. Use and maintain cell cultures. 29. Apply appropriate statistical and analytical techniques to test results. 30. Perform standard biochemical tests for organism identification. 31. Use specialized techniques for identification of bacteria and yeasts (e.g., fatty acids, electrophoresis, DNA probes, enzymelinked immunosorbent assay [ELISA], commercial kits). 32. Understand the advantages and limitations of various sterilization procedures. IV. LABORATORY OPERATIONS (24 questions) 33. Use appropriate safety techniques for the isolation and transfer of biological materials (e.g., loops, pipets, dilutor tips). 34. Handle, store, transport, and dispose of etiologic agents or biologics in compliance with laboratory and government regulations. 35. Handle, store, and dispose of hazardous chemicals and radiologic agents in compliance with laboratory and government regulations. 36. Document and maintain laboratory records and procedures. 37. Monitor the environment during process operations. 38. Maintain stock cultures. 39. Perform studies to determine sources of contamination.

11 40. Operate within environmentally controlled rooms, including clean rooms. 41. Evaluate clean-in-place and sterilize-in-place systems (e.g., validation procedures, monitoring procedures, troubleshooting). CP/QA Pharmaceutical/Medical Device/Cosmetic Specialty Tasks V. SAMPLE COLLECTION AND HANDLING (13 questions) 42. Prepare samples for microbiological analysis (e.g., sample size, blending, dilutions). 43. Collect and evaluate industrial samples for quality assurance and quality control testing. 44. Select appropriate methods for transport, handling, and storage of samples. VI. LABORATORY PROCEDURES (27 questions) 45. Isolate and identify yeasts of importance in industry. 46. Perform and validate tests for sterility. 47. Perform bacteriostatic or fungistatic tests. 48. Perform tests for particulate matter. 49. Perform and validate tests for pyrogens. 50. Perform and validate bioburden tests. 51. Perform tests for the effectiveness of preservatives. 52. Perform mutagen and cytotoxicity assays. 53. Perform process equipment and/or product validation studies. VII. STERILIZATION AND DEPYROGENATION VALIDATION (10 questions) CP/QA Food and Dairy Specialty Tasks VIII. SAMPLE COLLECTION AND HANDLING (12 questions) 59. Prepare samples for microbiological analysis (e.g., sample size, blending, dilutions, incubation conditions). 60. Collect and evaluate samples for environmental and quality control/quality assurance testing. 61. Select appropriate methods for transport, handling, and storage of samples. IX. LABORATORY PROCEDURES (38 questions) 62. Isolate and identify Listeria. 63. Isolate and identify Enterobacteriaceae (e.g., Salmonella, Proteus, Citrobacter, pathogenic Escherichia coli). 64. Isolate and identify Vibrio and Campylobacter. 65. Isolate and identify Clostridium species. 66. Isolate and identify microorganisms in food and dairy products. 67. Isolate, identify, and handle cultures of importance in food and dairy production (e.g., commercial starters). 68. Perform tests for spoilage and sterility in canned foods. 69. Perform shelf life studies. 70. Perform microbiological growth factor assays. 71. Perform tests for and identify extraneous materials in foods. 72. Use irradiation, biocontrol agents, preservatives, and other processing methods to control pathogens and spoilage bacteria in foods. 73. Perform aseptic process and/or product validation studies. 74. Perform direct microscopic counts. 54. Ethylene oxide sterilization. 55. Radiation sterilization. 56. Chemical and plasma sterilization. 57. Sterilization by filtration. 58. Depyrogenation. 11

12 CP/QA Sample Questions General CP/QA questions (for both Pharmaceutical/ Medical Device/Cosmetic and Food and Dairy examinations) 1. Phase-contrast microscopy enables the human eye to observe structures not visible by bright field-microscopy by modifying the light: a. path by 90. b. contrast. c. intensity. d. wavelength. e. amplitude. Corresponds to task #3. 2. To achieve Kohler illumination, the height adjustment of the condenser should be: a. at its upper stop. b. at its lower stop. c. halfway between its upper and lower stops. d. lowered slightly from its upper stop. Corresponds to task #3. 3. What is the greatest drawback in the use of UV-visible spectrophotometry in quantitative analysis? a. Inadequate linearity b. Inadequate sensitivity c. Inadequate specificity d. Excessive noise levels e. Problems in choosing appropriate blanks Corresponds to task #5. 4. Before a laminar flow hood is used, it should be running a minimum of: a. 2 hours. b. 24 hours. c. 15 to 30 minutes. d. 5 to 10 minutes. e. 1 week. Corresponds to task #7. 5. Which quality control procedure is necessary to perform with every batch of media? a. Shelf life determination b. Endotoxin content c. Buffering capacity d. Bacteriostatic/fungistatic tests e. Sterility check Corresponds to task # Pyrogallic acid with sodium hydroxide would be added for which of the following functions? a. Provide for anaerobic conditions b. Provide essential nutrients to the organisms c. Alter the ph of the medium d. Chelate metallic elements in the medium e. Bacteriocidal action Corresponds to task # When isolating motile, oxidative, nonfermenting gram-negative rods, which of the following should one look for in order to separate them from Pseudomonas species? a. An oxidative reaction on O/F glucose b. Polar pili c. The fermentation of dextrose d. Peritrichous versus polar flagella e. A negative indophenol oxidase reaction Corresponds to task #16. 12

13 8. A gram-negative organism was isolated with the following characteristics: oxidase-positive, motility-positive, growth at 42 C and production of pyocyanin. The organism isolated is: a. Escherichia coli. b. Pseudomonas cepacia. c. Pseudomonas aeruginosa. d. Staphylococcus aureus. e. Pseudomonas stutzeri. Corresponds to task # For the formation of immunoprecipitates, the almost universal optimal agar gel (Ouctherlony) concentration is: a to 0.15%. b to 0.3%. c. 0.3 to 1.5%. d. 1.5 to 3.0%. e. 3.0 to 5.0%. Corresponds to task # Precipitation tests for the detection of antigens are often carried out using: a. the agar diffusion method. b. high-performance liquid chromatography (HPLC). c. the complement fixation method. d. passive agglutination. e. polyacrylamide gel electrophoresis (PAGE). Corresponds to task # Fluorescence detected in immunofluorescent technique is photons emitted from the: a. antigen. b. antibody. c. antiglobulin antibody. d. antigen-antibody complex. e. fluorescent dye. Corresponds to task # A microscope outfitted with excitation, suppression, and heat filters is necessary to detect: a. peritrichous flagella. b. immunofluorescence. c. capsules. d. beta-hemolysis. e. fat soluble granules. Corresponds to task # Cross-reactivity in serologic reactions may lead to false-positive reactions and can be expected when: a. antigens are not in optimal proportions to the antibodies. b. there is no electrolyte in the system. c. several antigens are closely related. d. complement has not been inactivated. e. the dilutent is hypotonic. Corresponds to task # The scientific literature contains a number of references showing that antibiotic potency tests carried out with saturated paper disks are equivalent to the United States Pharmacopeia (USP) cylinder plate method. A laboratory wanting to change from cylinders to paper disks should: a. proceed to do so without further ado. b. provide definite proof of equivalence of the two tests for each antibiotic from the literature. c. provide proof of equivalence for each family of antibiotics for each medium used. d. provide some proof of equivalence for each test organism. e. provide some proof of equivalence for each antibiotic. Corresponds to task #21. 13

14 15. The agar diffusion test is the most convenient for antimicrobic susceptibility testing. However, antimicrobial dilution tests may be required if: a. simple qualitative information is needed. b. isolates are capable of growing at a uniform rapid rate. c. the drug has no diffusion problems. d. a fairly large number of drugs need to be screened at the same time. e. quantitative information is needed. Corresponds to task # The time at a given temperature required to destroy 90% of the organisms is called the: a. thermal death time. b. thermal death point. c. D value. d. F value. e. Z value. Corresponds to task # A 24-hour culture of Bacillus subtilis contains CFU/ml. Sequential dilutions of 1:10, 1:5, 1:100, and 1:3 were made from the original samples. The final titer is: a CFU/ml. b CFU/ml. c CFU/ml. d CFU/ml. e CFU/ml. Corresponds to task # The process of disinfection refers to: a. the destruction of disease-producing organisms. b. sterilization. c. the removal of all bacteria. d. the killing of all vegetative bacteria. e. the destruction of all bacterial spores. Corresponds to task # What standard is used for comparing the effectiveness of certain disinfectants? a. Iodine index b. Phenol coefficient c. Alcohol index d. Hexachlorophene coefficient e. Creosol index Corresponds to task # In order to identify a gram-negative, aerobic, nonfermenting, oxidase-negative bacterium, one should use a commercial kit which differentiates: a. genus Propionibacterium from Fusobacterium. b. genus Pseudomonas from Acinetobacter. c. genus Fusobacterium from Actinomyces. d. genus Fusobacterium from Bacteroides. e. various Enterobacteriaceae. Corresponds to task # Which of the following should be added to a commercial identification kit in order to create anaerobic conditions? a. Nitrogen gas b. Water c. Mineral oil d. Plastic wrap e. Sterile cotton plugs Corresponds to task # A laboratory's liability for its hazardous waste ends: a. when the hazardous waste is legally removed from the premises. b. when the hazardous waste is diluted and poured down the drain. c. when the waste has been mixed with hazardous wastes from another source by another party. d. when the waste no longer exists or is recycled. Corresponds to task #35. 14

15 23. The Resources Conversion and Recovery Act (RCRA) for hazardous waste requires: a. 60% of all medical waste to be recycled. b. medical waste to be disposed of within the state generated. c. a separate loading dock for hazardous waste. d. a cradle-to-grave tracking system. e. licensing requirements for class IV pathogens. Corresponds to task #35. Pharmaceutical/Medical Device/Cosmetics Specialty Questions 24. Which of the environmental conditions listed below would be selective of a mesophilic, anaerobic heterotroph? Temperature ph Atmosphere Energy Source a. 10 o C 7.0 N 2 glucose b. 60 o C 3.0 air sulfur c. 60 o C 1.0 air glucose d. 10 o C 7.0 air glucose e. 30 o C 7.0 N 2 glucose Corresponds to tasks #42 and According to Standard Methods for the Examination of Water and Wastewater, what is the maximum allowable time at 4 C for potable water samples that are to be plated? a. 6 hours b. 24 hours c. 30 hours d. 36 hours e. 48 hours Corresponds to task # The following units were sent to the laboratory for testing: 100 sterile samples for sterility tests 10 non-sterile samples for bioburden tests 10 non-sterile samples for pyrogen tests (in vitro) 3 extra sterile samples The units were packaged and shipped in a cardboard box. When the packages were opened with a razor box opener, two of the nonsterile packages were found to be damaged. How can the problem BEST be solved? a. Proceed by using the two damaged units for the bioburden test and note the damage on the report. b. Use two sterile units for the bioburden test. c. Use two sterile units for the pyrogen test. d. Perform the pyrogen test on eight units only, as sterility data will provide adequate information on pyrogen safety. e. Use two of the units for both the in vitro pyrogen test and the bioburden test. Corresponds to task # The growth of all bacteria requires that bacteriological liquid media provide an energy source plus: a. KH 2 PO 4, MgSO 4, (NH 4 ) 2 SO 4, and trace elements. b. vitamins, a buffer, trace elements, and sulfur. c. a buffer, O 2, vitamins, and a nitrogen source. d. hydrolytic products of proteins, vitamins and other growth factors, a buffer and trace elements. e. appropriate sources of C, H, N, P, and S, as well as trace elements. Corresponds to task #43. 15

16 28. One of the problems with the USP sterility test is that the test cannot be used to detect contamination in: a. ointments and oils. b. large numbers of contaminated containers from a lot of 10,000. c. small numbers of contaminated containers from a lot of 10,000. d. parenteral antibiotic solutions. e. monoclonal antibodies. Corresponds to task # What is the minimum detectable range of manual particle counts at 100, according to the USP? a. 5 µm b. 10 µm c. 25 µm d. 50 µm e. 100 µm Corresponds to task # Which indicates the MOST toxic cellular response in an in vitro cytotoxicity assay? a. Nonconfluent cell monolayer b. Granulation c. Crenation d. Acidic shift in ph of growth medium Corresponds to task #52. Food and Dairy Specialty Questions 32. The food-poisoning toxins produced by Staphylococcus aureus are: a. exotoxins. b. lethal poisons. c. endotoxins. d. heat labile. e. composed of carbohydrates. Corresponds to task # Saccharomyces cerevisiae var. ellipsoideus may be differentiated from Saccharomyces cerevisiae by: a. ph requirement. b. sugar fermentation. c. morphology. d. ascospore formation. e. amino acid requirements. Corresponds to task # Which of the following is a direct microscopic count technique? a. Smear b. Microcytic count c. Macrocytic count d. Spiral count e. Acid wash Corresponds to task # The two most common gases used for plasma sterilization are: a. hydrogen peroxide and peracetic acid. b. argon and hydrogen. c. hydrogen peroxide and ethylene oxide. d. hydrogen peroxide and acetic acid. Corresponds to task #56. 16

17 ANSWERS 1. a 6. a 11. e 16. c 21. c 26. c 31. a 2. d 7. d 12. b 17. e 22. d 27. e 32. a 3. c 8. c 13. c 18. a 23. d 28. c 33. c 4. c 9. c 14. e 19. b 24. e 29. b 34. a 5. e 10. a 15. a 20. b 25. c 30. a RESOURCES General Tasks Cunniff, P.A., et al. (ed.). Official methods of analyis. AOAC International, Gaithersburg, Md. Standard methods for the examination of water and wastewater. American Public Health Association (APHA), Washington, D.C. Biosafety in microbiological and biomedical laboratories. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, and National Institutes of Health. U.S. Government Printing Office, Washington, D.C. U.S. Food and Drug Administration. Current good manufacturing practice in manufacturing, processing, packing, or holding of drugs; general. Title 21, Code of Federal Regulations, section 210 (21 CFR 210). U.S. Government Printing Office, Washington, D.C. Pharmaceutical/Medical Device/Cosmetic Tasks Critical References United States Pharmacopeia and National Formulary. The United States Pharmacopeial Convention, Inc. Rockville, Md. Selected titles from the Association for Advancement of Medical Instrumentation (AAMI). Selected titles from Parenteral Drug Association Technical Monographs. Helpful References Journals PDA Journal of Pharmaceutical Science and Technology. Medical Device & Diagnostic Industry. The Microbiological Update. Food and Dairy Tasks Critical References Standard methods for the examination of dairy products. American Public Health Association, Washington, D.C. U.S. Food and Drug Administration (FDA) Bacteriological analytical manual (BAM), 8 th ed. AOAC International, Arlington, Va. Updates available at Vanderzant, C., and D.F. Splittstoesser (ed.). Compendium of methods for the microbiological examination of foods. American Public Health Association, Washington, D.C. 17

18 Helpful References Dey, B.P., and C.P. Lattuda (ed.) Microbiology laboratory handbook, 3 rd ed. United States Department of Agriculture, Food Safety and Inspection Service. U.S. Department of Agriculture, Washington, D.C. Journals Food Technology. Food Testing and Analysis. Journal of Food Protection. Journal of Validation Technology. The Microbiological Update. Internet USP. AAMI. AOAC. APHA. FDA. 18

19 Public Health and Medical Laboratory Microbiology (PH/MLM) Task List General Tasks I. LABORATORY INSTRUMENTS AND EQUIPMENT (19 questions) 1. Use and understand the principles of a steam autoclave and dryheat sterilizer. 2. Use of equipment which provides aerobic, anaerobic, capneic, and microaerophilic atmospheres. 3. Use and understand the principles of biological safety cabinets. 4. Use of bright-field, dark-field, fluorescence, stereoscopic, and phase contrast microscopy. 5. Use automated equipment for detection, identification, and/or antimicrobial susceptibility testing. 6. Use and understand the principles of immunoassay equipment. 7. Use and understand the principles of basic computer applications. II. LABORATORY PREPARATIONS (23 questions) 8. Use nonimmunologic stains for bacteria, fungi, parasites, and viruses. 9. Use diagnostic molecular microbiology techniques for the direct detection of infectious agents. 10. Use general, selective, and differential media for the isolation, identification, and preservation of bacteria. 11. Use general, selective, and differential media for the isolation, identification, and preservation of fungi. 12. Use cell culture techniques for the propagation of viruses and Chlamydia. 13. Use screening procedures to determine suitability of specimens for further microbiological analysis. 14. Use stains, wet mounts, reagents, equipment, media, and incubation conditions essential for specimen processing. III. SAMPLE COLLECTION AND HANDLING (16 questions) 15. Select appropriate specimens for microbiological analysis. 16. Select methods for collection and transport of specimens. 17. Ensure that all specimens are collected, transported, and analyzed in a safe manner. 18. Ensure proper preparation, packaging, and labeling of specimens sent to a reference laboratory. 19. Select criteria and procedures for rejection of inappropriate specimens. 20. Select appropriate criteria for specimen storage prior to analysis. 21. Ensure timely collection and transportation of specimens. IV. LABORATORY PROCEDURES (71 questions) 22. Isolate and identify significant anaerobic bacteria. 23. Isolate and identify significant gram-positive aerobic bacilli. 24. Isolate and identify mycobacteria. 25. Isolate and identify Enterobacteriaceae. 26. Isolate and identify glucose nonfermenting gram-negative bacteria. 27. Isolate and identify aerobic gram-negative cocci. 28. Isolate and identify clinically significant uncommon gramnegative bacilli (e.g., Bordetella, Brucella, Legionella, Pasteurella). 29. Isolate and identify Campylobacter, Vibrio, Aeromonas, Plesiomonas, and Helicobacter. 30. Isolate and identify Haemophilus. 31. Isolate and identify Staphylococcus and Micrococcus. 32. Isolate and identify Streptococcus and related microorganisms. 33. Isolate and identify microorganisms in water, food, and dairy products. 34. Isolate and identify the agents of superficial, cutaneous, and subcutaneous mycoses. 35. Isolate and identify opportunistic fungi. 19

20 36. Isolate and identify the agents of deep-seated mycoses. 37. Identify intestinal and urogenital protozoa. 38. Identify blood- and tissue-dwelling protozoa. 39. Identify intestinal and tissue-dwelling helminthes. 40. Isolate and identify Mycoplasma, Ureaplasma, and Chlamydia. 41. Isolate and/or identify the following viral agents: herpes simplex virus, varicella-zoster virus, cytomegalovirus (CMV), rotavirus, and respiratory syncytial virus. 42. Perform and interpret serologic tests for hepatitis A, hepatitis B, hepatitis C, and human immunodeficiency virus (HIV). 43. Use and understand the principles of the following assays for antibody detection: particle agglutination, enzyme immunoassays, and immunofluorescence. 44. Use serologic tests for the diagnosis of syphilis, Epstein-Barr virus (EBV), streptococcus, toxoplasma, rubella, CMV, and herpes simplex virus infections. 45. Perform and interpret antimicrobial susceptibility tests. 46. Perform assays for cytotoxin of Clostridium difficile. 47. Use and understand the principles of commercial kit identification systems. V. LABORATORY OPERATIONS (21 questions) 48. Laboratory safety, including biohazardous material, chemical and radioactive hazards, and fire safety. 49. Evaluate new products, develop new procedures, and modify existing procedures. 50. Laboratory administration: accreditation, proficiency testing, and workload. 51. Laboratory administration: fiscal analysis. 52. Laboratory administration: selecting personnel, evaluating laboratory staff, and overseeing continuing education (staff development). 53. Develop and maintain an effective quality assurance program. 54. Participate in epidemiological surveillance and infection control activities. 55. Consult with physicians and other laboratory users. PH/MLM Sample Questions 1. Oil immersion microscopy improves resolution by: a. increasing the numerical aperture. b. decreasing the numerical aperture. c. increasing the wavelength. d. decreasing the wavelength. e. increasing the magnification. Corresponds to task #4. 2. Utilization of chromogenic substrates for automated rapid bacterial identification systems depend on: a. bacterial growth. b. added reagents. c. oil overlay. d. cell wall antigens. e. bacterial enzyme activity. Corresponds to task #5. 3. False-positive growth indices may be obtained with automated blood culture systems due to: a. the presence of antimicrobial agents. b. an increased blood glucose level. c. an increased number of white blood cells. d. an insufficient volume of blood. e. a failure to increase the aeration of the aerobic vials by shaking. Corresponds to task #5. 20

21 4. Thiosulfate-citrate-bile-sucrose (TCBS) agar is used for the isolation of: a. Vibrio. b. Yersinia. c. Bordetella. d. Legionella. e. Haemophilus. Corresponds to task # Which of the following describes the optimum culture conditions for the recovery of Haemophilus ducreyi? a. 18 to 24 hours, 5 to 10% CO 2, 35 to 37 C b. 24 to 48 hours, 10 to 20% CO 2, 33 C c. 2 to 4 days, 5 to 10% CO 2, 35 to 37 C d. 5 to 7 days, 10 to 20% CO 2, 35 to 37 C e. 2 to 4 days, 5 to 10% CO 2, 33 to 35 C Corresponds to task # To determine the number of bacteria present in a tissue sample, 2.0g of ground tissue are suspended in sterile diluent so that the final volume is 10.0 ml. From this initial dilution, two successive 1:100 dilutions are made. Then, 1.0 ml of the final dilution is pipetted into a sterile petri dish to which is added 20.0 ml of sterile nutrient agar. The plate count after incubation is 54 colonies. The number of bacteria in 1.0 g of tissue is: a b c d e Corresponds to task # Polyvinyl alcohol (PVA)-preserved specimens can be kept without deterioration as long as several: a. minutes. b. hours. c. days. d. weeks. e. months. Corresponds to task # The patient is suspected of having a lung infection caused by Pneumocystis carinii. The specimen of choice treated with the methanimine silver stain that would MOST likely furnish a definite diagnosis is: a. an induced-sputum specimen. b. blood for antibody studies. c. a transtracheal aspirate. d. blood for antigen studies. e. a bronchial alveolar lavage. Corresponds to task # For short-term storage of specimens for viral culture, it is recommended that the sample be stored at: a. 20 C. b. 50 C. c. 70 C. d. 2 to 10 C. e. 22 to 25 C. Corresponds to task #20. 21

22 10. Urine specimens for culture should be transported to the laboratory immediately after collection and processed within 2 hours. An alternative for storage is to: a. freeze the specimen until it is processed, but not longer than 24 hours. b. place the specimen in the incubator for 8 hours. c. hold the specimen at room temperature for 8 hours. d. refrigerate the specimen until it is processed, but not longer than 24 hours. e. collect in a sterile container and store at room temperature for up to 4 hours. Corresponds to task # A catalase-positive, hippurate-positive, gram-positive organism exhibiting beta-hemolysis on blood agar and isolated from a case of neonatal sepsis could further be characterized by a positive: a. latex agglutination test. b. coagulase test. c. nitrate reduction. d. tumbling motility in wet mount. e. urease reaction. Corresponds to task # Nocardia brasiliensis can be differentiated from Streptomyces somaliensis by: a. casein decomposition. b. tyrosine decomposition. c. starch decomposition. d. acid from lactose. e. acid fastness. Corresponds to task # In order to isolate Erysipelothrix rhusiopathiae from skin lesions, the specimen should be inoculated into/onto: a. 1% glucose infusion broth. b. McBride agar. c. a Pai slant. d. clotted, heated rabbit blood. e. Skirrow medium. Corresponds to task # Mycobacterial pigmentation is due to: a. teichoic acid. b. lipid A. c. carbolic acid. d. carotenoids. e. mycolic acid. Corresponds to task # The test for indole production requires a culture to be inoculated into a medium rich in: a. lysine. b. ornithine. c. phenylalanine. d. pyruvate. e. tryptophan. Corresponds to task # Chromosomally mediated penicillin resistance in Neisseria gonorrhoeae: a. has not been seen in the United States. b. is detected with the chromogenic cephalosporin assay. c. is more prevalent than plasmid-mediated penicillin resistance. d. has been demonstrated in outbreaks of gonorrhea. e. produces penicillinase. Corresponds to task #27. 22

23 17. The identification of Bordetella pertussis from cultures is most commonly confirmed by: a. carbohydrate fermentation. b. microscopic characteristics. c. plasmid analysis. d. fluorescent-antibody testing. e. motility. Corresponds to task # Which of the following staphylococci is coagulase positive? a. S. haemolyticus b. S. hominis c. S. capitis d. S. intermedius e. S. saprophyticus Corresponds to task # The physiological property which is used to distinguish fecal coliforms is their ability to: a. hydrolyze esculin. b. resist bacteriophage infection. c. produce indole and be methyl red positive. d. grow at 44.5 C in appropriate media. e. hemagglutinate human red blood cells. Corresponds to task # Which of the following phrases describes a characteristic of hepatitis Be antigen (HBeAg)? a. Late indicator of acute active infection b. Usually long lived (3 to 6 months) c. Represents the most infectious period d. Useful in confirming chronic hepatitis B infection e. Persistence beyond the acute stage may indicate chronic liver disease and carrier state Corresponds to task # The total cell number of a bacterial broth culture can be determined directly by: a. carbohydrate consumption. b. electronic particle (cell) counts. c. most-probable-number method. d. plate counts. e. protein determination. Corresponds to task # A laboratory serving a hospital with 300 beds is currently processing all stool specimens using gram-negative enrichment broth, MacConkey agar, Hektoen enteric agar, and Campylobacter-selective blood agar. Which of the following media should be added on request for additional gram-negative enteric pathogens? a. MacConkey sorbital agar and cefsulodin-irgasan novobiocin (CIN) agar plates b. TCBS and bismuth-sulfite agar plates c. CIN and salmonella-shigella (SS) agar plates d. TCBS and xylose-lysine-desoxycholate (XLD) agar plates e. CIN and XLD agar plates Corresponds to task # Minor changes in a laboratory procedure are BEST made by a: a. memo to all employees. b. change or addition to the original procedure, dated and signed. c. complete version of the entire procedure. d. verbal explanation of the change at a laboratory meeting. e. verbal explanation to the person who performs the test. Corresponds to task #50. 23

24 24. A laboratory is classified as an extent 4 laboratory in mycobacteriology by the College of American Pathologists. Which of the following is the appropriate response from a laboratory at this extent level? a. Mycobacterium tuberculosis; photochromogen; Mycobacterium fortuitum b. Mycobacterium tuberculosis; photochromogen; rapid grower c. Mycobacterium tuberculosis; two mycobacteria other than Mycobacterium tuberculosis d. Mycobacterium tuberculosis; Mycobacterium kansasii; rapid grower e. Mycobacterium tuberculosis; Mycobacterium kansasii; Mycobacterium fortuitum Corresponds to task # A technical procedure must always include the test principle, which is: a. a brief statement concerning the type of reaction(s) involved and the reasons for performing the test. b. an outline of steps involved in collection procedures. c. a list of reagents, supplies, and equipment. d. a brief statement concerning tolerance limits for controls. e. a brief statement concerning limitations of the procedures. Corresponds to task # A workload reporting system is an important part of laboratory management because it: a. tells exactly how much should be charged per test. b. keeps personnel busy. c. counts only tests done and specimens received in the laboratory without inflating these figures by adding quality control and standardization efforts. d. helps in planning, developing, and maintaining efficient laboratory services with administrative and budget controls. e. complies with regulatory requirements. Corresponds to task # Which step must be taken when a laboratory reports an incorrect identification on a proficiency test sample? a. No corrective action is required. b. Document both the error and the corrective action taken. c. Review testing policy and procedure. d. Request a follow-up specimen. e. Provide an inservice to review the problem with all employees. Corresponds to task # According to most laboratory certifying agencies, procedure manuals should be reviewed by supervisory personnel at least: a. monthly. b. quarterly. c. semiannually. d. annually. e. biannually. Corresponds to task # Which of the following situations would most likely cause a drop in the paid productivity of a department from 45 minutes/hour in June to 35 minutes/hour in July? a. Decreased use of vacation time in July b. Replacement of a 4-hour identification procedure with a 2- hour procedure starting July 1 c. Increased number of routine tests in July d. Training a new technologist in July e. Addition of a new laboratory testing procedure in July Corresponds to task # To determine the specificity of a test, one must: a. multiply the positive predictive value by 100. b. divide true positives by (true positives plus false negatives). c. divide true negatives by (false positives plus true negatives). d. divide true positives by total positives. e. subtract false positives from total positives. Corresponds to task #54. 24

25 31. Which of the following specimens would you recommend to a physician who must monitor a burn victim for the presence of invasive infection? a. Quantitative burn eschar b. Blood cultures c. Gram-stained smears of burn sites d. Histopathological examination of tissue e. Routine burn cultures Corresponds to task # The physician should always be notified by telephone when a: a. nares culture is positive. b. stool culture is positive. c. blood culture is positive. d. cerebral spinal fluid (CSF) culture is negative. e. gonorrhea culture is positive. Corresponds to task #55. ANSWERS 1. a 6. c 11. d 16. d 21. b 26. d 31. b 2. e 7. a 12. e 17. d 22. a 27. c 32. c 3. c 8. c 13. a 18. d 23. b 28. d 4. a 9. d 14. d 19. d 24. c 29. d 5. e 10. d 15. e 20. c 25. a 30. c RESOURCES Critical References Forbes, B. A., D. F. Sahm, and A. S. Weissfeld (ed.) Bailey & Scott s diagnostic microbiology, 10 th ed. Mosby, Inc., St. Louis, Mo. Isenberg, H. D. (ed.) Clinical microbiology procedures handbook. American Society for Microbiology, Washington, D.C. Murray, P. R., E. J. Baron, J. H. Jorgensen, M. A. Pfaller, and R. H. Yolken (ed.) Manual of clinical microbiology, 8 th ed. ASM Press, Washington, D.C. Helpful References Persing, D.H., T.F. Smith, F.C. Tenover, and T.J. White (ed.) Diagnostic molecular microbiology: principles and applications. ASM Press, Washington, D.C. Rose, N.R., R.G. Hamilton, and B. Detrick (ed.) Manual of clinical laboratory immunology, 6 th ed. ASM Press, Washington, D.C. Journal Diagnostic Microbiology and Infectious Disease. Internet ASM. Centers for Disease Control and Prevention (CDC). 25

26 Consumer and Industrial Microbiology (C & I) Task List General Tasks I. LABORATORY INSTRUMENTS AND EQUIPMENT (19 questions) 1. Use and monitor sterilization equipment (e.g., autoclaves, ovens, ethylene oxide, radiation). 2. Use and monitor filters for sterilization of solutions. 3. Use and monitor incubation devices such as ambient air, carbon dioxide, anaerobic, and constant water temperature devices. 4. Use a ph meter or conductivity meter. 5. Use locally controlled environmental systems (e.g., biosafety cabinets, unidirectional [laminar] flow cabinets, isolators). 6. Use various types of microscopes. 7. Use colorimetric and spectrophotometric equipment. 8. Use of electronic monitoring of equipment (chart recorders, multipoint recorders) and understand electronic data trending (storage, retrieval, auditing, electronic signature; Code of Federal Regulations, title 21, section 11 [21 CFR 11]). 9. Calibrate and maintain laboratory equipment. II. LABORATORY PREPARATIONS (9 questions) 10. Evaluate media (general, selective, and differential) for growth, isolation, and identification of bacteria and fungi. 11. Perform medium growth promotion tests. 12. Use stains, both general and for specific structures (spores, flagella, capsules). 13. Prepare solutions of known molarity, molality, and normality. III. SAMPLE COLLECTION AND HANDLING (2 questions) 14. Select appropriate means of disposal for analyzed or unanalyzed samples. 15. Use appropriate documentation procedures for samples involved in legal actions (e.g., chain-of-custody documentation). IV. LABORATORY PROCEDURES (28 questions) 16. Understand and apply biochemical tests for bacterial identification (e.g., carbohydrate fermentations, redox reactions, catalase, coagulase, oxidase). 17. Isolate and identify coliforms. 18. Isolate and identify Pseudomonas species and common waterborne organisms. 19. Isolate and identify common fungi (e.g., Aspergillus species, yeast). 20. Perform analytical procedures for the evaluation of water and potable water. 21. Perform measurements for the growth of microorganisms (e.g., substrate utilization, plate counts, turbidity). 22. Use most-probable-number technique. 23. Perform microbial tests on disinfectants. 24. Perform identification of bacteria using biochemical, genetic, or chromatographic procedures (e.g., DNA probes, polymerase chain reaction [PCR], sequencing, fatty acid methyl esters, carbohydrate utilization). 25. Understand rapid microbiological techniques (e.g., bioluminescence, impedence, cytometry). 26. Use or detect viruses and/or mycoplasma. 27. Evaluate new test procedures or procedures that are alternative to compendial procedures. 26