S IMPLE AM PLIFICATION B ASED - A SSAY
|
|
|
- David Riley
- 6 years ago
- Views:
Transcription
1 S IMPLE AM PLIFICATION B ASED - A SSAY a point of care platform for resource-limited settngs Helen Lee & team Diagnostics for the Real World University of Cambridge
2 Complexity of automated NAT - Abbott Real time PCR Room 1 - Sample processing and extraction
3 Abbott HIV RealTime PCR system Room 2 Detection
4 Disposables & reagents for 1,000 Roche PCRtests Dry Ice
5 Reducing complexity of NAT 1. Converting complicated detection system into a visual signal Reference Wicking Pad Target sequence Capture Zone Capture probe Detection Complex Dineva et al, JCM 2007
6 3. Simplifying front end sample preparation SAMBA Roche Qiagen Storage at 4 o C No Yes Yes Alcohol No Yes Yes Chaotropic salts No Yes Yes Preparation of reagents by users No Unit dose Yes Carrier RNA & ethanol Yes Carrier RNA & ethanol
7 5. Develop stable reagents to avoid cold chain transport freeze balls reagents Sample Preparation Amplification Enzyme 1 Internal Control Reagent 1 Enzyme 2 Reagent 2
8 % Signal Strength Real time stability data of amplification enzymes C 4 C 25 C 37 C Now 12 months Time Shipping stability - stable at 55 C for 1 month
9 4. Closed amplification cartridge containing all liquid & dry reagents Prevent contamination of amplified products
10 Roche added target located in LTR but only in some assays principally used in developed world 5 LTR Gag Vif LTR 3 Pol Env Nef SAMBA BioMerieux Roche v. 1 Abbott Sequence variation: Env > Gag > Pol > LTR
11 Signal strength SAMBA detection of HIV-1 WHO subtype panel A B C D AE F G AGGH 712 (CRF02) NIBSC standard (B) Group N Group O Negative plasma Group M (400 copies/ml) SAMBA primers detect all subtypes, including groups N and O
12 Blinded evaluation of SAMBA performance with NIH external quality assurance subtype panels (Rush University Medical Center, J Bremer) Subtype N pos / N tested A 12/12 A/E 42/42 A/G 9/9 C 36/36 D 18/18 F 30/30 G 12/12 TOTAL 168/168 (100%)
13 SAMBA system for HIV diagnosis Visual detection of nucleic acids on a dipstick Unit dose device containing pre-loaded reagents Simplified sample preparation Integrated internal control Results < 90 minutes Qualitative test (Plasma or whole blood) Acute infection during window period Early infant diagnosis Semi-quantitative copies/ml (plasma) Viral level for therapy monitoring
14 SAMBA 1 semi-automated instruments
15 SAMBA test visual readout NEGATIVE LOW POSITIVE
16 Blinded comparison of SAMBA qualitative assay (plasma) vs. Roche Taqman v1 in HIV patients at Royal London Hospital Subtype No. samples Virus level No. SAMBA + A B C D F G H J K x x x10 5 7x x x x x10 3-2x10 5 1x10 3-2x /31 24/25 30/30 9/9 7/7 5/5 1/1 4/4 5/5 Overall Detection rate: 116/117 (99.1%)
17 SAMBA detection of HIV recombinant subtypes vs. Roche Taqman v.1.0 (Royal London Hospital) Subtype No. samples Viral level No. SAMBA + CRF01_AE CRF02_AG CRF06_CPX CRF13_CPX CRF14_BG AD A/CRF02_AE Other CRF Untyped x x10 6 8x10 3-2x x10 3-3x10 4 1x10 3-2x x10 5 3x10 4-4x10 7 2x10 4-5x10 5 6/6 19/20 2/2 1/1 5/5 11/11 9/9 15/15 5/5 Overall detection rate: 73/74 (98.6%)
18 Roche v1 negative HIV samples detected by SAMBA (Barts hospital, London) Subtype SAMBA signal Roche result Abbott HIV copies/ml C 5 0 1,118 A/D 1 0 1,780 K 5 0 1,028 CRF02_AG ,276 CRF02_AG ,508 CRF13_CPX
19 Performance of SAMBA detection of HIV-1 clinical samples (Barts hospital, London) Sample Number No. SAMBA + HIV positives /191 HIV patients With undetectable viral load (<40 copies/ml) 9 0/9 HIV negative 216 0/216 SAMBA sensitivity (Roche + Abbott combined) = 98 % (189/191) 0 false positive in 225 clinical samples
20 Roche added target located in LTR but only in some assays principally used in developed world 5 LTR Gag Vif LTR 3 Pol Env Nef BioMerieux Roche v. 1 Abbott SAMBA Roche Taqman v. 2 Sequence variation: Env > Gag > Pol > LTR
21 SAMBA performance vs. Roche Taqman No. samples Clade Sensitivity Specificity 485 (plasma) A, B, C, D, F, G, H, J, K, recombinants 99.2 % (252/254) 100 % (231/231) 144 (whole blood) ND 98 % (48/49) 100 % (95/95) % (300/303) 100% (326/326) *Roche Taqman v1.0 (Royal London Hospital) for 416 samples; Roche Taqman v2 for subsequent 213 samples
22 Major Challenges of EID LTFU between testing and treatment Country LTFU Kenya 83% Mozambique 69% - 87% Uganda 79% Malawi 77% Swaziland 82% Zambia 37% Clinton Foundation
23 Blind evaluation of SAMBA EID with NIH HIV-1 DNA sensitivity EQA Panel (Rush University Medical Center, J Bremer) HIV DNA copies/test SAMBA Roche Amplicor v /6 0/6 8 7/7 1/7 (5/7 indeterminate) 20 6/7 6/7 40 5/5 5/5 80 5/5 5/5 TOTAL 29/30 23/30
24 Evaluation of SAMBA EID test in whole blood from adult patients attending HIV clinic (London Barts Hospital) Whole blood SAMBA tested blinded DBS prepared on-site Frozen aliquot Zambia (Amplicor) Uganda (TaqMan) Royal London Hospital Proviral DNA PCR using multiple genes
25 Roche Roche SAMBA EID test vs. Roche DNA Amplicor v1.5 by DBS (Lusaka, Zambia) INITIAL SAMBA + - RETEST SAMBA * 50-9* 50 *14/22 DBS false-negative became positive, 1 positive became negative SAMBA (whole blood) Roche (DBS) Initial Retest Sensitivity 100 % (114/114) 81 % (92/114) 92 % (105/114)
26 Roche Roche SAMBA EID test vs. Roche Ampliprep/TaqMan by DBS (Kampala, Uganda) INITIAL RETEST SAMBA + - SAMBA * 50 *11/16 DBS false-negatives became positive, 3 positives became negative Sensitivity SAMBA (whole blood) 100 % (114/114) Initial 86 % (98/114) Roche (DBS) Retest 93 % (106/114)
27 Field evaluation of SAMBA semi-quantitative assay at Chiradzulu, Malawi 200 patients recruited by MSF staff from HIV clinic during routine visit 200 µl fresh plasma SAMBA by MSF staff on-site Data sent blinded Frozen plasma shipped directly Roche TaqMan v2 Royal London Hospital Discordant Samples Abbott RealTime PCR Addenbrooke s Hospital All data sent to MSF for analysis
28 Roche Roche Blinded validation of SAMBA semi-quant in 134 clinical samples St Thomas & Royal London Hospitals In absolute counts SAMBA <1000 >1000 < * Within 0.3 Log accuracy of Roche SAMBA <1000 >1000 < >1000 3** 34 > *2/4 within 0.3 Log accuracy of Roche ** 2/3 within 0.3 Log accuracy of Roche Concordance between SAMBA & Roche Taqman v % (131/134)
29 Malawi results SAMBA vs Roche TaqMan v2 Viral Load (cp/ml) < 500 (<1.7 Log) Roche TaqMan VL ( Log)* >2,000 >(3.3 Log) Total SAMBA >1, SAMBA < 1, Total (%) 146 (73%) 8 (4%) 46 (323%) 200 * Roche TaqMan is 0.3 Log accuracy (package insert) Overall concordance with Roche v2 = 98% (196/200)
30 Uganda results SAMBA vs Roche TaqMan v2 Viral Load (cp/ml) < 500 (<1.7 Log) Roche TaqMan VL * ( Log) >2,000 >(3.3 Log) Total SAMBA >1, SAMBA < 1, Total (%) 92 (60%) 2 (1%) 60 (39%) 154 * Roche TaqMan is 0.3 Log accuracy (package insert) Overall concordance with Roche v2 = 96.1% (148/154)
31 Conclusion SAMBA platform is much simpler than currently available molecular technologies which require highly-trained personnel and sophisticated infrastructure only available in centralised laboratories SAMBA device is much easier to handle and being a closed system, prevents contamination by amplicons Staff training requirement for SAMBA is minimal SAMBA can be implemented in lower healthcare levels such as district hospitals or health centres with a basic laboratory but supplied with electricity Reagents do not need cold chain transport or cold storage, a considerable advantage in resource-poor settings
32 Dilution series of whole blood clinical samples for blinded evaluation of SAMBA & DBS in Zambia & Uganda Plasma VL (cp/ml) ARV start Last CD4 Subtype 2 samples with high VL ,021 No 412 A , B 2 samples with low VL * G C 2 samples with undetectable VL B C
33 Limit of detection: SAMBA, TaqMan Qual Test and Amplicor v1.5 (dilutions of 2 samples with high viral load) DNA (cp/ml) RNA (cp/ml) SAMBA TaqMan Amplicor Sample 1547, VL 269,021, subtype A 3, , / , /+ 34 2, /+ -/ /+ +/ /+ -/ / Sample 1346 VL 230,788 subtype B 3, , / , /+ 33 2, /+ -/ /+ +/ /- -/ /
34 Sensitivity of SAMBA, TaqMan Qual Test and Amplicor v1.5 using dilutions of 2 samples with low viral load DNA (cp/ml) RNA (cp/ml) SAMBA TaqMan Amplicor /+ +/ /- -/- Sample 1524 VL208/ml subtype G /- -/- -/ / / DNA (cp/ml) RNA (cp/ml) SAMBA TaqMan Amplicor Sample 1554, VL 83, subtype C /+ +/ /+ -/+ +/ /- -/- -/ /- - -
35 Sensitivity of SAMBA, TaqMan Qual Test and Amplicor v1.5 using dilutions of 2 samples with undetectable viral load DNA (cp/ml) RNA (cp/ml) SAMBA TaqMan Amplicor Sample 1348, VL 0, subtype B /+ +/ /+ -/+ +/ /- -/- -/ /- - - DNA (cp/ml) RNA (cp/ml) SAMBA TaqMan Amplicor Sample 1352, VL 0, subtype C /- +/ /+ +/- +/ /- -/+ -/ /- - -
36 Sensitivity of SAMBA, TaqMan Qual Test and Amplicor v1.5 using WHO 2 nd International RNA standard diluted in whole blood DNA (cp/ml) RNA (cp/ml) SAMBA Qual Test TaqMan Qual Test Amplicor DNA PCR v , , /+ -/ /- +/ /
37 Summary of SAMBA, TaqMan Qual Test and Amplicor v1.5 sensitivity in dilutions of clinical samples SAMBA Qual Test TaqMan Qual Test Amplicor DNA PCR v1.5 Sample type Whole blood DBS DBS Nucleic acid detection RNA/DNA RNA/DNA DNA Limit of detection in clinical samples (RNA cp/ml) Limit of detection In NIBSC RNA standard (IU/ml) Limit of detection In clinical samples (DNA cp/ml) NA NA
38 Sensitivity of SAMBA, TaqMan Qual Test and Amplicor v1.5 using 8E5 LAV cell line diluted in whole blood DNA (cp/ml) RNA (cp/ml) SAMBA Qual Test TaqMan Qual Test Amplicor DNA PCR v , ,000,000 +/ ,000 16,500,000 +/ ,000 1,650,000 +/+ + +/ ,000 +/+ + +/ ,500 +/+ + -/ / ,650 +/ /+ +/ /+ +/ /+ -/ Amplicor only detects DNA; SAMBA & TaqMan detect both RNA & DNA
39 Comparison of SAMBA with Roche TaqMan v.2 in dilution series (100 ul whole blood) Roche TaqMan v.2 RNA cp/ml plasma Number SAMBA Qualitative Test In whole blood HIV negative blood donors (Ab neg, Roche Amplicor neg) 95 0 HIV positive RNA negative patients 23 22* 40 cp/ml * Pos on repeat testing 49 48
40 Next steps: EID clinical trials Cameroon Protocol in development Judith Shang CDC - Cameroon Kenya Protocol submitted, awaiting approval Clement Zeh CDC - Kenya & KEMRI Malawi Protocol approved T. Banda-Kuthyola MOH South Africa Protocol in final stage of preparation Gayle Sherman - NHLS Ashraf Coovadia - MRC & RMMC Hospital Uganda Protocol approved by local IRB; awaiting approval by US IRB Mary Glenn Fowler - Makarere & Johns Hopkins Adeodata Kikitiinwa - Baylor-Uganda Clinic Zambia Protocol submitted Charles Nyambe - MOH Jeff Stringer - CIDRZ Zimbabwe Protocol approved Sekesai Zinyowere - MOH
41 SAMBA 2 instrument prototype
42 ISO 13485:2003 and CAN/CSA ISO certified CA-FDA Licensed medical device manufacturing facility
43 Dry room <5% humidity
44 Vanuatu prevalence: 23 % - Validated bottled drinking water - Sustainable implementation a bigger challenge!
45 Analysis of 8 repeat DBS false negatives at Zambia by Roche Amplicor v1.5 ID Roche TaqMan VL Clade SAMBA (blood) Zambia (manual Amplicor v1.5) Uganda (automated Ampliprep/ TaqMan) Initial Repeat Initial Repeat Proviral DNA (blood) 97 2,979 C ND ND ,450 ND CRF02_AG <40 ND D C
Alere q. A platform to answer global health needs: TB and beyond. Duncan Blair Director, Public Health Initiatives
Alere q A platform to answer global health needs: TB and beyond Duncan Blair Director, Public Health Initiatives 7 th FIND Symposium, Barcelona, October 2014 Alere q platform overview Comprising the Alere
Instrument specifications Technology Software Host interfaces Printer interfaces Data Station Physical dimensions Weight Certifications
The evolution of PCR Introducing the COBAS TaqMan 48 Analyzer At the forefront of PCR evolution Since Roche acquired the rights to PCR in 1991, its mission has been to fully realize the potential of this
Instructions for Use. RealStar Lassa Virus RT-PCR Kit /2017 EN
Instructions for Use RealStar Lassa Virus RT-PCR Kit 1.0 04/2017 EN RealStar Lassa Virus RT-PCR Kit 1.0 For research use only! (RUO) 641003 INS-641000-EN-S01 96 04 2017 altona Diagnostics GmbH Mörkenstr.
1001 NW Technology Drive, Lee's Summit, MO // (800) // (816) Fax //
Herpes Simplex Virus 1 & 2 (HSV-1 & HSV-2) Quantitative Real-time PCR Test Code: 8500 Some specimen types for this assay are reported as qualitative results; please see our Specimen Information section
Product# TM Intended Use. use only and not for use in diagnostic procedures.
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Norovirus TaqMan RT-PCR Kit Product# TM41400 Product Insert Intended
HELINI White spot Syndrome virus [WSSV] Real-time PCR Kit
![HELINI White spot Syndrome virus [WSSV] Real-time PCR Kit](/thumbs/72/67570787.jpg "HELINI White spot Syndrome virus [WSSV] Real-time PCR Kit")
HELINI White spot Syndrome virus [WSSV] Real-time PCR Kit Instruction manual Cat. No: 6001-25/50/100 tests Compatible with: Agilent, Bio-Rad, Roche, Applied Bio systems [ABI], Rotor-gene, Cepheid, Bioer,
BEIPH Final Report. EQA Programme 2011 Hepatitis B Virus (HBVDNA11A) William G Mackay on behalf of QCMD and its Scientific Council July 2011
BEIPH Final Report EQA Programme 2011 Hepatitis B Virus (HBVDNA11A) William G Mackay on behalf of QCMD and its Scientific Council July 2011 Not to be reproduced or quoted without permission of QCMD. Any
Positioning of TB Diagnostics within a Tiered System Integrated Approach from Reference to District Laboratory. Giorgio Roscigno CEO FIND
Positioning of TB Diagnostics within a Tiered System Integrated Approach from Reference to District Laboratory Giorgio Roscigno CEO FIND Integrated Laboratory Network Definition An integrated laboratory
Best Practices for the Use of International Standards in Vaccine Testing Dianna Wilkinson
Best Practices for the Use of International Standards in Vaccine Testing Dianna Wilkinson DCVMN Regional Training Workshop Hyderabad, 07-10 May, 2018 Biologicals substances which cannot be fully characterized
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: (905) Fax: (905)
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com HPV (High Risk) TaqMan PCR Kit Product# TM32200 Product Insert
Intended Use Norgen s Parvovirus B19 TaqMan PCR Kit is designed for the detection of Parvovirus B19
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Parvovirus B19 TaqMan PCR Kit Product# TM39600 Product Insert
Use of MagNA Pure 96 System and LightCycler Instrument for Testing of Nail Samples for Dermatophytes Detection
MagNA Pure System Application Note No. 7 March 2015 Use of MagNA Pure 96 System and LightCycler Instrument for Testing of Nail Samples for Dermatophytes Detection Eveline Snelders 1, Arjan S. de Jong 1,
Varicella Zoster Virus Real Time PCR Kit
Revision No.: ZJ0001 Issue Date: Aug 28th, 2007 Varicella Zoster Virus Real Time PCR Kit Cat. No.: OD-0024-01 For use with LightCycler2.0/ LightCycler480 (Roche) real time PCR systems For in vitro Diagnostic
Molecular Diagnostics for Global Heath Problems. Proactive Investors Forum 5 October 2017
Molecular Diagnostics for Global Heath Problems Proactive Investors Forum 5 October 2017 2 Document Information The information contained in this document and made verbally to you (together the Presentation
CASE-STUDY- VALIDATION of PCR based methodology. Beata Surmacz-Cordle Senior Analytical Development Scientist
CASE-STUDY- VALIDATION of PCR based methodology Beata Surmacz-Cordle Senior Analytical Development Scientist UK RMP Pluripotent Stem Cell Platform Validation Workshop 2 nd June 2016 RT-qPCR assay for detection
How to Choose the Right Equipment/Platforms for your Laboratory
How to Choose the Right Equipment/Platforms for your Laboratory (A Public Hospital Laboratory Perspective) Michelle J. Francis Overview Public Hospital Pressures / Challenges Commercial vs In-house assays
MAXimum results. Minimum effort.
MAXimum results. Minimum effort. MagMAX sample preparation for veterinary diagnostic testing Sometimes purifying nucleic acid from animal samples can be like trying to pull a needle out of a haystack.
LightCycler 480 qpcr Tools. Meeting the Challenge of Your Research
LightCycler 480 qpcr Tools Meeting the Challenge of Your Research Find the Optimal LightCycler 480 Reagents for Your Research Application: Are you analyzing DNA DNA Nucleic acid isolation Manual processing
Novel Diagnostic Methods of Tuberculosis
Novel Diagnostic Methods of Tuberculosis Qimin YOU USTAR Biotechnologies I. The TB Diagnosis Challenge My presentation will focus on the new diagnostic methods of tuberculosis, developed by USTAR Biotechnologies.
Intended Use Norgen s Giardia intestinalis End-Point RT-PCR Kit is designed for the detection of Giardia
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Giardia intestinalis End-Point RT-PCR Kit Product# EP43800 Product
Quality Assurance at the Point of Care AM Manufacturer's Perspective. Eugen Ermantraut Alere Technologies, Jena
Quality Assurance at the Point of Care AM Manufacturer's Perspective Eugen Ermantraut Alere Technologies, Jena 1 Definitions POC Testing: Diagnostic testing ti that t is performed near to or at the site
Dengue Virus subtypes 1,2 3 and 4
TM Primerdesign Ltd Dengue Virus subtypes 1,2 3 and 4 3 Untranslated Region (3 UTR) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Dengue Virus subtypes 1,2
MOLECULAR TESTING: VERIFYING/VALIDATING INSTRUMENTS, REAGENTS AND ASSAYS. Richard L. Hodinka, Ph.D.
MOLECULAR TESTING: VERIFYING/VALIDATING INSTRUMENTS, REAGENTS AND ASSAYS Richard L. Hodinka, Ph.D. University of South Carolina School of Medicine Greenville Greenville Health System, Greenville, SC hodinka@greenvillemed.sc.edu
Herpes Simplex 1 IgM (HSV1 IgM)
DIAGNOSTIC AUTOMATION, INC. 23961 Craftsman Road, Suite D/E/F, Calabasas, CA 91302 Tel: (818) 591-3030 Fax: (818) 591-8383 onestep@rapidtest.com technicalsupport@rapidtest.com www.rapidtest.com See external
Transfusion-transmitted Virus(TTV) Real Time PCR Kit
Revision No.: ZJ0003 Issue Date: Aug 7 th, 2008 Transfusion-transmitted Virus(TTV) Real Time PCR Kit Cat. No.: HD-0013-02 For use with ABI Prism 7000/7300/7500/7900; Smart CyclerII; icycler iq 4/iQ 5;
Molecular Diagnostics
Molecular Diagnostics Part II: Regulations, Markets & Companies By Prof. K. K. Jain MD, FRACS, FFPM Jain PharmaBiotech Basel, Switzerland May 2018 A Jain PharmaBiotech Report A U T H O R ' S B I O G R
Mycobacterium tuberculosis End-Point PCR Kit Product# EP42100
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Mycobacterium tuberculosis End-Point PCR Kit Product# EP42100
Update on Pediatric diagnostics, EID and POC
UNICEF / Supply Division Health Technology Centre (HTC) Medical Unit Update on Pediatric diagnostics, EID and POC Ludo Scheerlinck Technical Officer, Medical Devices & Clinical Laboratory UNICEF Supply
Trichomonas vaginalis SYBR Green PCR Kit Product# SG52000
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Trichomonas vaginalis SYBR Green PCR Kit Product# SG52000 Product
MAXimum results. Minimum effort.
MAXimum results. Minimum effort. MagMAX sample preparation for veterinary diagnostic testing Sometimes purifying nucleic acid from animal samples can be like trying to pull a needle out of a haystack.
Development of a Second Version of the Cobas AmpliPrep/Cobas TaqMan Hepatitis C Virus Quantitative Test with Improved Genotype Inclusivity
JOURNAL OF CLINICAL MICROBIOLOGY, Sept. 2011, p. 3309 3315 Vol. 49, No. 9 0095-1137/11/$12.00 doi:10.1128/jcm.00602-11 Copyright 2011, American Society for Microbiology. All Rights Reserved. Development
Plasma/Serum RNA/DNA Purification Mini Kit Product# 55200
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Plasma/Serum RNA/DNA Purification Mini Kit Product# 55200 Product
Bill Kabat and Carmen Irizarry of the ACTG/IMPAACT Lab Tech Committee. Network Name, Title Signature Date
Title: Dried PBMC Pellet Processing Standard Operating Procedure (LDMS specimen code = BLD/EDTA or ACD/PEL) Origination Date: 01 Jun 2004 Total Pages: 6 Effective Date: 29 Nov 2012 SOP Number LTC-SOP-59
Real-Time PCR Workshop Gene Expression. Applications Absolute and Relative Quantitation
Real-Time PCR Workshop Gene Expression Applications Absolute and Relative Quantitation Absolute Quantitation Easy to understand the data, difficult to develop/qualify the standards Relative Quantitation
Roche Molecular Biochemicals Technical Note No. LC 10/2000
Roche Molecular Biochemicals Technical Note No. LC 10/2000 LightCycler Overview of LightCycler Quantification Methods 1. General Introduction Introduction Content Definitions This Technical Note will introduce
Dengue Virus subtypes 1, 2, 3 and 4 (Multiplex kit)
Primerdesign TM Ltd Dengue Virus subtypes 1, 2, 3 and 4 (Multiplex kit) genesig advanced kit 100 tests For general laboratory and research use only 1 Contents Introduction to Dengue Virus 3 Specificity
Stool GeneXpert MTB/Rif Assay
Stool GeneXpert MTB/Rif Assay Standard Operating Procedure 1.0. Purpose The purpose of this standard operating procedure (SOP) is to detail the steps for correctly performing, interpreting, and documenting
Targeted Sequencing in the NBS Laboratory
Targeted Sequencing in the NBS Laboratory Christopher Greene, PhD Newborn Screening and Molecular Biology Branch Division of Laboratory Sciences Gene Sequencing in Public Health Newborn Screening February
Epstein Barr Virus (Human Herpes virus 4)
TM Primerdesign Ltd Epstein Barr Virus (Human Herpes virus 4) nonglycosylated membrane protein (BNRF1) gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to
Isolation of Viral RNA, Viral DNA and Bacterial DNA from Animal Samples
Application Note Isolation of Viral RNA, Viral DNA and Bacterial DNA from Animal Samples Denis Flüge, Lillian Krüger, Sandy Leifholz and Holger Engel R&D Department, QIAGEN GmbH, QIAGEN Straße 1, Hilden,
Human T-lymphotropic Virus 1
TM Primerdesign Ltd Human T-lymphotropic Virus 1 Polymerase gene (POL) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human T-lymphotropic Virus 1 Human T-lymphotropic
Human T-lymphotropic Virus 1
PCRmax Ltd TM qpcr test Human T-lymphotropic Virus 1 Polymerase gene (POL) 150 tests For general laboratory and research use only 1 Introduction to Human T-lymphotropic Virus 1 Human T-lymphotropic Virus
Instructions for Use. RealStar HEV RT-PCR Kit /2017 EN
Instructions for Use RealStar HEV RT-PCR Kit 2.0 07/2017 EN RealStar HEV RT-PCR Kit 2.0 For use with Mx 3005P QPCR System (Stratagene) VERSANT kpcr Molecular System AD (Siemens Healthcare) ABI Prism 7500
Principals of Real-Time PCR. Amira A. T. AL-Hosary Lecturer of Infectious Diseases, Faculty of Veterinary Medicine, Assiut University, Egypt
Principals of Real-Time PCR Amira A. T. AL-Hosary Lecturer of Infectious Diseases, Faculty of Veterinary Medicine, Assiut University, Egypt What Is Real-Time PCR? Nucleic acid (DNA) amplification and detection
Viral Hemorrhagic Septicemia Virus
Techne qpcr test Viral Hemorrhagic Septicemia Virus Non-coding region 150 tests For general laboratory and research use only 1 Introduction to Viral Hemorrhagic Septicemia Virus Viral Hemorrhagic Septicaemia
Product# EP Product Description
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com Chlamydia End-Point PCR Kit Product# EP31400 Product Insert Intended
Own the future As molecular testing evolves, so can you
Own the future As molecular testing evolves, so can you Not for distribution in the US; not available in all markets. cobas 8800 System Introducing the cobas 6800 and 8800 Systems Redefining molecular
Enteroinvasive E. Coil(EIEC) Real Time PCR Kit. For In Vitro Diagnostic Use Only User Manual
Revision No.: ZJ0003 Issue Date: Aug 7 th, 2008 Enteroinvasive E. Coil(EIEC) Real Time PCR Kit Cat. No.: DD-0093-01 For Use with LightCycler 1.0/LightCycler2.0/LightCycler480 (Roche) Real Time PCR Systems
IgG Antibodies To Toxoplasma. Gondii ELISA Kit
IgG Antibodies To Toxoplasma Catalog No: IRAPKT1405 Gondii ELISA Kit Lot No: SAMPLE INTENDED USE The Toxoplasma IgG ELISA is intended for use in evaluating a patient s serologic status to Toxoplasma gondii
! "! " # $ % & ' $ (!)* + * ",- '(!)* +. '/0 $ - 1 $
!"!" #$%& ' $ (!)*+*",-'(!)*+.'/0$- 1 $ + 234 1506417 5" 8/" (!)*+ 9 '(!)" 9 ' 9 '(!)*+* ' 9 1 / 1" '" 8' " '2 " 8',, 5+ 234 15064 17 5(!)"7, " 2 " '2 ' " 6 64 17 "," " " 64 1 "22 : " " " 641 2,0 '," )
Human Papillomavirus 16
TM Primerdesign Ltd Human Papillomavirus 16 E6 gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human Papillomavirus 16 Papillomaviruses are a diverse
Real-time RT-PCR Protocol for the Detection of A(H7N9) Influenza Virus
Real-time RT-PCR Protocol for the Detection of A(H7N9) Influenza Virus 8 April 2013 The WHO Collaborating Center for Reference and Research on Influenza at the, Beijing, China, has made available attached
Acinetobacter baumannii
TM Primerdesign Ltd Acinetobacter baumannii hypothetical protein sequence gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Acinetobacter baumannii Acinetobacter
Acinetobacter baumannii
TM Primerdesign Ltd Acinetobacter baumannii hypothetical protein sequence gene genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Acinetobacter baumannii Acinetobacter
Dedicated to Molecular Diagnostics
Dedicated to Molecular Diagnostics For Europe Sample & Assay Technologies QIAGEN s growing role in molecular diagnostics At QIAGEN, one of our key goals is to support clinical decision-making and improve
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: (905) Fax: (905)
3430 Schmon Parkway Thorold, ON, Canada L2V 4Y6 Phone: 866-667-4362 (905) 227-8848 Fax: (905) 227-1061 Email: techsupport@norgenbiotek.com E.coli O157:H7 End-Point PCR Kit Product# EP41300 Product Insert
QUALITY CONTROL/QUALITY ASSURANCE IN THE MOLECULAR MICROBIOLOGY LABORATORY
QUALITY CONTROL/QUALITY ASSURANCE IN THE MOLECULAR MICROBIOLOGY LABORATORY Richard L. Hodinka, Ph.D. University of South Carolina School of Medicine Greenville Greenville Health System, Greenville, SC
SYBR Green Realtime PCR Master Mix
Instruction manual SYBR Green Realtime PCR Master Mix 0810 F0924K SYBR Green Realtime PCR Master Mix QPK-201T 1 ml x 1 QPK-201 1 ml x 5 Contents [1] Introduction [2] Components [3] Primer design [4] Detection
The Seed Health Toolbox II. Molecular Seed Testing Methods and Processes: What are the sources of variability?
The Seed Health Toolbox II Molecular Seed Testing Methods and Processes: What are the sources of variability? Stephan C. Brière Manager Plant Pathology Diagnostic Laboratory, Ottawa, ON Canadian Food Inspection
Toxoplasma gondii Repeat region. genesig Advanced Kit. 150 tests. Primerdesign Ltd. For general laboratory and research use only
TM Primerdesign Ltd Toxoplasma gondii Repeat region genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Toxoplasma gondii Toxoplasma gondii is a species of parasitic
Human Herpes Virus 1 (Herpes simplex type 1)
TM Primerdesign Ltd Human Herpes Virus 1 (Herpes simplex type 1) Capsid assembly and DNA maturation gene genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Human
Human Herpes Virus 1 (Herpes simplex type 1)
TM Primerdesign Ltd Human Herpes Virus 1 (Herpes simplex type 1) Capsid assembly and DNA maturation gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human
TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0
Cat. # 9766 For Research Use TaKaRa MiniBEST Viral RNA/DNA Extraction Kit Ver.5.0 Product Manual Table of Contents I. Description... 3 II. Components... 3 III. Storage and shipping... 3 IV. Preparation
Functional Genomics Research Stream. Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update
Functional Genomics Research Stream Research Meeting: June 19, 2012 SYBR Green qpcr, Research Update Updates Alternate Lab Meeting Fridays 11:30-1:00 WEL 4.224 Welcome to attend either one Lab Log thanks
SideStep Lysis and Stabilization Buffer
SideStep Lysis and Stabilization Buffer INSTRUCTION MANUAL Catalog #400900 Revision B.0 For Research Use Only. Not for use in diagnostic procedures. 400900-12 LIMITED PRODUCT WARRANTY This warranty limits
Selection and use of Ebola in vitro diagnostic (IVD) assays
EMERGENCY GUIDANCE Selection and use of Ebola in vitro diagnostic (IVD) assays June 2015 World Health Organization 2015. All rights reserved. The designations employed and the presentation of the material
Convoy TM Transfection Reagent
Convoy TM Transfection Reagent Catalog No.11103 0.25ml (40-80 transfections in 35mm dishes) Catalog No.11105 0.5 ml (80-165 transfections in 35mm dishes) Catalog No.11110 1.0 ml (165-330 transfections
Human T-lymphotropic Virus 1
TM Primerdesign Ltd Human T-lymphotropic Virus 1 Polymerase gene (POL) genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Human T-lymphotropic Virus 1 Human T-lymphotropic
Fast decisions instead of delays Quality Control through outstanding design
Fast decisions instead of delays Quality Control through outstanding design We are CustomBiotech from Roche In your operations, behind your decisions, powering your products You are driving a paradigm
PAXgene Blood DNA PAXgene Blood DNA Tube (IVD) for clinical use PAXgene Blood DNA System (RUO) for research use. Explore more at
PAXgene Blood DNA PAXgene Blood DNA Tube (IVD) for clinical use PAXgene Blood DNA System (RUO) for research use Explore more at www.preanalytix.com Situation The composition, amount, quality and integrity
Quantitation of mrna Using Real-Time Reverse Transcription PCR (RT-PCR)
Quantitation of mrna Using Real-Time Reverse Transcription PCR (RT-PCR) Quantitative Real-Time RT-PCR Versus RT-PCR In Real-Time RT- PCR, DNA amplification monitored at each cycle but RT-PCR measures the
Technical Review. Real time PCR
Technical Review Real time PCR Normal PCR: Analyze with agarose gel Normal PCR vs Real time PCR Real-time PCR, also known as quantitative PCR (qpcr) or kinetic PCR Key feature: Used to amplify and simultaneously
CaMV promoter & NOS terminator detection
Primerdesign TM Ltd GMO event quantification CaMV promoter & NOS terminator detection Detection and quantification of GMO integration events in Soya by real-time PCR 100 tests Kit contents CaMV-GM primer/probe
Human Herpes Virus 6. genesig Standard Kit. BHLF3 (putative dutpase gene) 150 tests. Primerdesign Ltd. For general laboratory and research use only
TM Primerdesign Ltd Human Herpes Virus 6 BHLF3 (putative dutpase gene) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Human Herpes Virus 6 HHV-6B is responsible
Hepatitis A Virus. genesig Standard Kit 5 NCR. 150 tests. Primerdesign Ltd. For general laboratory and research use only
TM Primerdesign Ltd Hepatitis A Virus 5 NCR genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Hepatitis A Virus Hepatitis virus (HAV) is a non-enveloped ssrna
Acinetobacter baumannii
PCRmax Ltd TM qpcr test Acinetobacter baumannii hypothetical protein sequence gene 150 tests For general laboratory and research use only 1 Introduction to Acinetobacter baumannii Acinetobacter baumannii
Human Papillomavirus 16
TM Primerdesign Ltd Human Papillomavirus 16 E6 gene genesig Advanced Kit 150 tests For general laboratory and research use only 1 Introduction to Human Papillomavirus 16 Papillomaviruses are a diverse
Foot & Mouth Disease Virus
TM Primerdesign Ltd Foot & Mouth Disease Virus 5' untranslated region (5'UTR) genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Foot & Mouth Disease Virus Foot-and-mouth
AmoyDx TM JAK2 V617F Mutation Detection Kit
AmoyDx TM JAK2 V617F Mutation Detection Kit Detection of V617F mutation in the JAK2 oncogene Instructions For Use Instructions Version: B1.0 Date of Revision: July 2012 Store at -20±2 o C -1/5- Background
Executive Summary. clinical supply services
clinical supply services case study Development and NDA-level validation of quantitative polymerase chain reaction (qpcr) procedure for detection and quantification of residual E.coli genomic DNA Executive
Received 3 December 2008/Returned for modification 17 February 2009/Accepted 14 July 2009
JOURNAL OF CLINICAL MICROBIOLOGY, Sept. 2009, p. 2872 2878 Vol. 47, No. 9 0095-1137/09/$08.00 0 doi:10.1128/jcm.02329-08 Copyright 2009, American Society for Microbiology. All Rights Reserved. Evaluation
AmoyDx JAK2 Mutation Detection Kit
AmoyDx JAK2 Mutation Detection Kit Detection of V617F mutation in the JAK2 oncogene Instructions For Use For Research Use Only and For Reference Only Instructions Version: B1.2 Date of Revision: June 2016
Quantifiler Human DNA Quantification Kit Quantifiler Y Human Male DNA Quantification Kit
Product Bulletin Human Identification Quantifiler Human DNA Quantification Kit Quantifiler Y Human Male DNA Quantification Kit The Quantifiler kits produce reliable and reproducible results, helping to
In the name of God. Application of Molecular Methods for Detection of Viral Pathogens & Viral Resistance
In the name of God Application of Molecular Methods for Detection of Viral Pathogens & Viral Resistance By: Kiana Shahzamani From :Digestive Disease Research Center (DDRC) Why detect microorganisms? Determine
UltraFast Molecular Diagnostic System
UltraFast Molecular Diagnostic System CONTENTS 01 PCR vs Real-time PCR 02 NANOBIOSYS Sample Prep G2-16TU 03 NANOBIOSYS Real-time PCR G2-4 01 PCR vs Real-time PCR What is DNA & What is PCR? NANOBIOSYS 4
Transmissible gastroenteritis virus
TM Primerdesign Ltd Transmissible gastroenteritis virus Spike Protein (S) Gene genesig Standard Kit 150 tests For general laboratory and research use only 1 Introduction to Transmissible gastroenteritis
Relative Quantification (Mono-Color) Unknown samples (purified total RNA, mrna or cdna or genomic DNA)
The LightCycler 480 System Short Guide Topic: Purpose: Assay Principle: Detection Format: Result: Relative Quantification (Mono-Color) Describes how to set up and perform mono-color Relative Quantification
RealLine HPV 31 / 33 Str-Format
Instructions for use A QUALITATIVE ASSAY KIT FOR THE DIFFERENTIAL DETERMINATION OF HUMAN PAPILLOMAVIRUS TYPE 31 AND 33 DNA BY REAL TIME PCR METHOD In vitro diagnosticum () VBD8471 96 Tests valid from May
FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE.
Instruction manual RNA-direct SYBR Green Realtime PCR Master Mix 0810 F0930K RNA-direct SYBR Green Realtime PCR Master Mix Contents QRT-201T QRT-201 0.5mLx2 0.5mLx5 Store at -20 C, protected from light
TOSCANA VIRUS oligomix Alert kit reagent for cdna "nested" amplification
TABLE OF CONTENTS INTENDED USE page 1 KIT DESCRIPTION page 1 KIT CHARACTERISTICS page 2 OTHER PRODUCTS REQUIRED page 2 MATERIALS PROVIDED IN THE KIT page 2 MATERIALS REQUIRED BUT NOT PROVIDED IN THE KIT
GENOMERA TM MRSA/SA PRODUCTS A NEW ERA IN DIRECT MRSA DNA TESTING. Bringing genetic MRSA results in less than 1 hour!
GENOMERA TM MRSA/SA PRODUCTS A NEW ERA IN DIRECT MRSA DNA TESTING Bringing genetic MRSA results in less than 1 hour! IMPROVING PATIENT CARE THROUGH EARLIER DETECTION OF MRSA AND SA FAST MRSA AND SA DETECTION
American Society of Cytopathology Core Curriculum in Molecular Biology
Brought American Society of Cytopathology Core Curriculum in Molecular Biology Brought American Society of Cytopathology Core Curriculum in Molecular Biology Chapter 3 Molecular Techniques Alternatives
virotype ASFV PCR Kit Handbook
virotype ASFV PCR Kit Handbook August 2015 24 (catalog no. 281903) 96 (catalog no. 281905) For detection of DNA from African swine fever virus (ASFV) Licensed in accordance with 11 (2) of the German Animal
validation report for TaqMan Roundup Ready Quantification Kit
validation report for TaqMan Roundup Ready Quantification Kit Life Technologies offers a breadth of products DNA RNA protein cell culture instruments For Research Use Only. Not intended for any animal
ReverTra Ace qpcr RT Master Mix
Instruction manual ReverTra Ace qpcr RT Master Mix 1203 F1173K ReverTra Ace qpcr RT Master Mix FSQ-201 200 reactions Store at -20 C Contents [1] Introduction [2] Components [3] Protocol 1. RNA template
RealLine Food Kit Turkey Detect
Instruction for Use DETECTION KIT FOR DNA FROM ALL KIND OF TURKEY IN EXTREMELY PROCESSED AND CANNED FOOD AND IN OTHER PRODUCTS BY REAL-TIME-PCR This Kit is for Food Diagnostic only! CIB50700 100 tests
Developing an Accurate and Precise Companion Diagnostic Assay for Targeted Therapies in DLBCL
Developing an Accurate and Precise Companion Diagnostic Assay for Targeted Therapies in DLBCL James Storhoff, Ph.D. Senior Manager, Diagnostic Test Development World Cdx, Boston, Sep. 10th Molecules That
AmpliSens Enterovirus-EPh PCR kit
For Professional Use Only AmpliSens Enterovirus-EPh PCR kit Instruction Manual AmpliSens Ecoli s.r.o., Studenohorska 12 841 03 Bratislava 47 Slovak Republic Tel.: +421 2 6478 9336 Fax: +421 2 6478 9040
Acute bee paralysis virus
Techne qpcr test Acute bee paralysis virus Replicase polyprotein 150 tests For general laboratory and research use only 1 Introduction to Acute bee paralysis virus Since its first identification, the presence
Genomic DNA quantification assay. Quantification of genomic DNA by real-time PCR
Genomic DNA quantification assay Quantification of genomic DNA by real-time PCR Contents Kit contents 3 Reagents and equipment to be supplied by user 3 Kit storage and stability 4 Suitable sample material