Difficulty to implement An Integrated Molecular Biology tool in Latin America (exemplified by Brazil)

Transcription

1 UFBA Serviço de Gastro Hepatologia e Laboratório de Pesquisa em Infectologia Difficulty to implement An Integrated Molecular Biology tool in Latin America (exemplified by Brazil) by Andreas Stocker (MD) astocker@ufba.br

2 Diagnostics of viral Hepatitis Detection Treatment Serological Immunoassays For qualitative Determination of Antibodies Selection Genotyping: PCR and Sequencing or Hybridization Monitoring Viral Load: Real-time PCR Evaluation of Mutations: PCR and Sequencing or Hybridization

3 Laboratory Conditions I Serological Analysis Closed System 1 Room Up to analysis per month Price per analysis: R$ 18,- ($6,50)

4 Laboratory Conditions II PCR Laboratory of 3 (4) separated Areas Geografical Separation Storage + Nucleic Acids Extraction Geografical Separation Pre- PCR Room X Preparing of PCR tubes + Plates X PCR- and Post PCR Area

5 Laboratory Conditions III Closed Systems for Viral Load Analysis Closed System 1 Room Up to analysis per month Price per analysis: R$ 500,- ($210,-)

6 Laboratory Centers Collection Point Collection Point Collection Point Collection Point Collection Point Laboratory Center Collection Point Collection Point

7 Blood Collection Sampling Conditions I Serological Analysis Centrifugation (Serum) (Aliquotation) Storage 25 C Storage 4 C Up to 4 hours (Preventation of Hemolysis) Up to 1 day Up to 1 week Storage -20 C

8 Sampling Conditions II Molecular Diagnostics Blood Collection Centrifugation (Serum/Plasma) Aliquotation Processing time Maximum: 1 hour! Storage -80 C 60

9 Sampling Conditions III Transport Conditions Serology Not frozen: -Up to 2 days at 4 C with Coolers -20 C Frozen: -In dry Ice at -20 C Molecular Diagnostics Frozen: -Up to 24h with Coolers -80 C -In dry Ice at -80 C

10 Transport of Specimen I

11 Transport of Specimen II

12 Coolers -80ºC Transport of Specimen III Never use Coolers -20ºC for Molecular Diagnostics Specimen!

13 Transport of Specimen IV Rigidious entrance control and protocolation of specimen s temperature -80ºC -70ºC -60ºC -50ºC -40ºC -30ºC -20ºC -10ºC -0ºC

14 Discrepancy of Results I Viral Load [LOG] 9,00 Viral Load A-Probe: Commercial System B-Probe: In House q-pcr 8,00 7,00 6,00 5,00 4,00 SGH/LAPI Acre 3,00 2,00 1,00 0,

15 Discrepancy of Results II Viral Load A- Probe: Comercial System B-Probe: In House q-pcr 7,00 6,00 5,00 Viral Load[LOG] 4,00 3,00 SGH/LAPI LOG10(CV) LACEN LOG10(CV) 2,00 1,00 0,00 H-053 H-059 H-064 H-067 H-108 H-109 H-124 H-128 H-135

16 Monitoring of the commercial Systems

17 Problems - Education level of the involved Technicians - Impossibility of blinded tests - Lack of recording of Sample Processes and Transport - Absence of a Monitoring System - Small amount of Personnel Resources

18 Solutions - Individual scheme for every collecting point - Special classes for technicians (obligatory) - Implementation of a Recording System - Implementation of a Monitoring System - Increase of Personnel Resources

19 Example I University Hospital Salvador/Bahia Quantity: HBV/HCV a month Laboratory: Central Laboratory (LACEN/Salvador) Transport: weekly; Coolers -80 C; 4hours

20 Example II Clinics Hospital (Fundhacre), Rio Branco/Acre Quantity: HBV/HCV a month Laboratory: Central Laboratory (LACEN/Rio Branco) Transport: - Explanation: Lack of collection area, infrastructure and technicians Solution: 5 times a week collection directly at the LACEN

21 Example III Dermatological Hospital, Cruzeiro do Sul/Acre Quantity: 100 HBV/HCV a month Laboratory: Central Laboratory (LACEN/Rio Branco) Distance: 600km Transport: by plane; Coolers -80 C; time: 12 hours Temporary suspended Lack of the grant for the technician

22 Example IV Specialized Assistance, Sena Madureira/Acre Quantity: 100 HBV/HCV a month Laboratory: Central Laboratory (LACEN/Rio Branco) Distance: 150km Transport: - Explanation: Lack of collection area, infrastructure and technicians Solution: Once a week bus transport of 25 patients to the Central Laboratory

23 Solutions - Individual scheme for every collecting point - Special classes for technicians (obligatory) - Implementation of a Recording System - Implementation of a Monitoring System - Increase of Personnel Resources

24 The Future I New Laboratory of Molecular Diagnostics Rio Branco

25 The Future II -Production of in-house PCR Kits licensed by ANVISA -Creation of small and simples real-time PCR Laboratories Pre-PCR Extraction PCR

26 UFBA Serviço de Gastro Hepatologia e Laboratório de Pesquisa em Infectologia Thank you very much for your attention! Any questions? by Andreas Stocker (MD) astocker@ufba.br