Applying technical requirements of ISO/IEC 17025:2005 in the Zoonotic diseases testing laboratory. Discussion on the challenges.

Transcription

1 Applying technical requirements of ISO/IEC 17025:2005 in the Zoonotic diseases testing laboratory. Discussion on the challenges. Mrs Masabata Annah Motaung: Agricultural Research Council - Onderstepoort Veterinary Research, Private Bag X5, Onderstepoort,, Gauteng, 0110, South Africa; Tel: ; Fax: ; MacholoA@arc.agric.za Introduction Laboratory facilities are designated as basic Biosafety Level (BSL) 1, basic Biosafety Level 2, containment Biosafety Level 3, and maximum containment Biosafety Level 4. Biosafety level designations are based on a composite of the design features, construction, containment facilities, equipment, practices and operational procedures required for working with agents from the various risk groups. Biological Safety Levels are a series of protections relegated to the activities that take place in particular biological labs. They are individual safeguards designed to protect laboratory personal, as well as the surrounding environment and community. These levels, which are ranked from one to four, are selected based on the agents or organisms that are being researched or worked on in any given laboratory setting. For example, a basic labs setting specializing in the research of non-lethal agents that pose a minimal potential threat to lab workers and the environment are generally considered BSL 1 (e.g. less virulent strains of Escherichia coli) the lowest biosafety lab level. A specialized research laboratory that deals with potentially deadly infectious agents like prions causing Bovine Spongiform Encephalopathy (BSE) disease would be designated as BSL3 the highest and most stringent level (Standard BS :1992) Bovine Spongiform Encephalopathy (BSE), which is commonly known as mad cow disease, is according to South African animal disease act of 1984, a controlled disease because it is zoonotic and has an economic impact ( Cattle are the primary hosts and are infected through eating rations that including scrapie-infected material (Chesebro, 1999). The disease can be transmitted to humans by ingestion of undercooked contaminated beef such as brain and spinal cord (Tylor et al, 1997,). The symptoms of the disease in cattle include an excitable or nervous temperament to Page 1 of 10

2 external stimuli such as touch to the skin and the inability to stand. In humans, it causes memory loss, emotional instability including inappropriate outbursts, unsteady gait, severe rapidly dementia and death. There is no treatment to cure this disease and neither there is an effective vaccine so the control is through test and slaughter. Infected animals are slaughtered and destroyed by incineration (Brown, 2003) Escherichia coli (E. coli) is a type of bacteria that normally live in the intestines of people and animals. However, some types of E. coli can cause intestinal infection. Symptoms of intestinal infection include diarrhoea, abdominal pain, and fever. More severe cases can lead to bloody diarrhoea, dehydration, or even kidney failure. People with weakened immune systems, pregnant women, young children, and older adults are at increased risk for developing these complications. Most intestinal infections are caused by contaminated food or water. Proper food preparation and good hygiene can greatly decrease chances of developing an intestinal infection. Most cases of intestinal E. coli infection can be treated at home. Symptoms generally resolve within a few days to a week (Bonadio et al, 2001) South Africa has been declared free of BSE however DAFF is monitoring BSE and the Agricultural Research Council is a reference laboratory to confirm this status. South African National Accreditation system (SANAS) is an accreditation body which asses technical and management activities as per ISO requirements. SANAS was given a task by DAFF to assess and recommend all controlled and notifiable diseases testing laboratories for accreditation ( ultimate goal was to improve the standard of all laboratories involved in diagnostic testing for controlled and notifiable diseases enabling the laboratories to obtain SANAS accreditation based on ISO In this paper we highlight the benefits and challenges associated with introduction, implementation and maintenance of internationally recognised quality standards (ISO 17025) in laboratories testing for zoonotic diseases. BSL 3 laboratory was compared Page 2 of 10

3 to BSL 1 laboratory with respect to technical requirements such as personnel training and competency, test method validation, proficiency testing, equipment calibration and maintenance Discussions Knowing the difference in biosafety laboratory levels and their corresponding safety requirements is imperative for anyone working with microbes in a laboratory setting. A summary of summary of biosafety levels for infectious agents is presented in Table 1. This table shows the differences between the biosafety level facilities, practices, primary barriers and safety equipment to protect personnel and prevent contamination Concerning PPE it has been observed that the personnel protective equipment (PPE) used in BSL 1 is comfortable and easy to wear i.e. normal laboratory coats and gloves. The test is performed on an open bench with or without a mask. However, personnel working in BSL 3 should wear a special PPE i.e. full suit, which covers the neck, arms, and legs, goggles and special mask to prevent infections. The environmental conditions should be controlled and monitored and access to the laboratory restricted to unauthorised personnel (untrained). Medical surveillance for personnel working in BSL 3 should be monitored regularly to ensure their safety. More than one personnel at a time should be inside the facility in case of reporting accidents and emergencies Personnel training and competency Personnel working in BSL 1 only need to practice basic good laboratory practice (GLP) and basic biosafety/ biosecurity measures and may enter the laboratory at any time any day and the access may be controlled by locking or code system. Whereas, for personnel working in BSL 3 needs to be properly trained on bio-risk aspects like, entrance and exit of the laboratory, access lights upon entering, showering, and the access control shall be traceable e.g. code system, finger print and surveillance camera before the actual testing therefore this may take time to declare personnel competent. Page 3 of 10

4 Test method validation and proficiency testing Validation of the BSL 1 test method can be done as per the TR57 because both positive and negative samples can be accessed. All validation parameters can be established, justified and the test method can easily be proven fit for purpose. Contrary, validation test BSL 3 methods may be very challenging because the country is free from the disease. Samples tested during the validation are all negative and this may lead to non-compliance with the TR57 because the validation parameters may not be established. The laboratory depend on the validation from the supplier of the kit used for testing BSE and Proficiency testing (PT) samples (positive and negative samples received). When PT samples are received, high biosafety precautions should be taken to ensure that the disease is not spread. In addition, other logistics like, application for import permit from DAFF, proper maintenance of the testing facility for containment of the disease, and SOP on handling of the samples upon receipt and proper disposal of the samples after testing are required Equipment calibration and maintenance When the equipment are be calibrated and maintained in BSL 3 laboratory the laboratory may be required to shut down for 1 to 2 weeks for major maintenance, repairs and equipment replacement. This shutdown will allow for other scheduled activities including annual re-certification for BSL 3 compliance, directional airflow, pressure decay verification and air exchange rates in conjunction with High Efficiency Particulate Air (HEPA) filter integrity testing, verification of the Building Management System (BMS) and the air-handling units. This can be challenging because the equipment calibration and maintenance schedule should be aligned with the annual shut down, customer communication to ensure customer satisfaction and that the turn-around time is not affected and compliance with ISO Equipment calibration and maintenance in BSL1 can be done as per schedule, on site and anytime. 125 Page 4 of 10

5 Document control Document control is one of the most important requirement in ensuring the consistency of practices, activities, processes and systems; clear transmission of information, minimum loss of information, improved understanding and elimination of uncertainty and confusion, provision of direction. In BSL1 facility, personnel may be able to generate, access documents and print them for reading at work and home. However, in BSL 4, the documents can be read electronically and should be contained in the laboratory and not be taken home to avoid contamination. In case of SANAS assessment documentation (audit checklists) shall be filled in electronically and other documents will be scanned and ed to another computer outside the facility for printing Conclusion It is evident that there are differences between testing for diseases classified as BSL 1 and BSL 3 in a laboratory setting. The working conditions, procedures, handling of test items, method validation, equipment calibration and maintenance, biosafetybiosecurity procedures, declaration of personnel competence, participating in Proficiency testing and document control are regarded as strenuous and challenging to implement in BSL 3 due to costs and risks. However, it is feasible to comply with ISO requirements in these laboratories. The procedures should be properly followed to avoid infections and to ensure containment of the diseases, personnel to be properly trained, and regular medical surveillance monitoring to effectively ensure and assure quality of results produced without compromising personnel s health. It is recommended that personnel are well compensated for discomfort to ease the burden posed by the biosecurity and biosafety requirements References AMERICAN BIOLOGICAL SAFETY ASSOCIATION Risk Group Classifications for Infectious Agents [online]. Available at Page 5 of 10

6 BONADIO M, MEINI M, SPETALERI P, GILGI C. Current microbiological and clinical aspects of urinary tract infections. Eur J Urol 2001; 40: Brown P. (2003). Transmissible spongiform encephalopathy as a zoonotic disease, International life sciences institute Chesebro, B. (1999). Prion protein and the transmissible spongiform encephalopathies. Neuron, 24: GUIDELINES FOR THE EVALUATION OF VETERINARY SERVICES. OIE TERRESTRIAL ANIMAL HEALTH CODE HARTMAN, A. L., HOMER, L. C., TRICHEL, A. M., FISHER, D. FREROTTE, J., & COLE, K. S Evolution of a Facility-specific BSL-3 Training Program for the University of Pittsburgh Regional Bio-containment Laboratory: Journal of Applied Biosafety Vol. 15, No. 3, University of Pittsburgh, Pittsburgh: Pennsylvania INTERNATIONAL ORGANIZATION FOR STANDARDAZATION (ISO) 17025:2005. Quality management systems - General requirements for the competence of testing and calibration laboratories. MICROBIOLOGICAL SAFETY CABINETS. Recommendations for selection, use and maintenance. London, British Standards Institution, 1992 (Standard BS :1992) OFFICE INTERNATIONAL DES EPIZOOTIES (OIE) QUALITY STANDARD AND GUIDELINES FOR VETERINARY LABORATORIES: INFECTIOUS DISEASES SANDIA NATIONAL LABORATORIES Laboratory Biosecurity and Biosafety for BSL3 Laboratories. India. SEWEL, C.L Laboratory-associated infections and biosafety. Clinical Microbiology Reviews. Vol8. no3: Page 6 of 10

7 SOUTH AFRICA Animal Diseases Act, No.35 of 1984 [online]. Available from: [Assessed: 07/03/2014]. SOP: DAFF 001. Standard for DAFF Approval of Veterinary Laboratories/ BSL3 Facility Compliance SOP: DAFF 002. Requirements for DAFF Approval/ Compliance 2013 SUTER II, G.W Risk Characterization for Ecological Risk Assessment of Contaminated Sites. Lockheed Martin Energy Systems, Inc. Oak Ridge, Tennessee Taylor DM et al. The effect of formic acid on BSE and scrapie infectivity in fixed and unfixed brain-tissue. Veterinary Microbiology, 1997, 58: WORLD HEALTH ORGANISATION Guidance on regulations for the transport of infectious substances. WHO/CDS/EPR/ WILSON, D.E., MEMARZADER, F.THEODORE, J. T. & RIDENHOUR, W.B Biosafety Level 3 Laboratory Certification Requirements. National Institutes of Health. New York: Van Nostrand. Page 7 of 10

8 212 Table 1. Summary of Recommended Biosafety Levels for infectious agents BSL level Agents Practices Primary barriers and safety equipment Facilities 1 Not known to consistently cause diseases in healthy adults. Standard microbiological practices No primary barriers required. PPE: laboratory coats and gloves, eye, face protection, as needed Laboratory bench top and sink 2 Agents associated with human BSL-I practice plus: limited access. Primary barriers; BSCs or other BSL-I plus Autoclave available disease. Routes of biohazard warning signs. sharps physical containment devices used transmission include precautions. Biosafety manual for all manipulations of agents that percutaneous injury, ingestion, defining any needed waste cause splashes or aerosols of mucous membrane exposure decontamination or medical infectious materials. PPE; Laboratory surveillance policies. coats, gloves, face and eye protection, as needed. Page 8 of 10

9 3 Indigenous or exotic agents BSL-II practice plus: controlled Primary barriers: BSL-II plus: that may cause serious or potentially lethal disease through the inhalation route of exposure. access. Decontamination of all waste. Decontamination of laboratory clothing before laundering. BSCs or other physical containment devices used for all open manipulations of agents. PPE: Protective laboratory clothing, gloves, face, eye and respiratory protection, as needed. Physical separation from access corridors. Self-closing, double-door access. Exhausted air not recirculated. Negative airflow into laboratory. Entry through airlock or anteroom. Hand washing sink near laboratory exit. 4 Dangerous/exotic agents BSL-III practices plus: Clothing Primary barriers: BSL-III plus: which post high individual risk of aerosol-trans-mitted laboratory infections that are frequently fatal, for which there are no vaccines or treatments. Agents with a close or identical antigenic relationship change before entering. Shower on exit. All material decontaminated on exit from facility. All procedures conducted in Class III BSCs in combination with full-body, air-supplied, positive pressure suit. Separate building or isolated zone. Dedicated supply and exhaust vacuum, and decontamination systems. Other requirements outlined in the text. to an agent requiring BSL-4 until data is available to redesignate the level. Related agents with unknown risk of transmission. Page 9 of 10

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