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Premium member Presentation Transcript CD4 for monitoring HIV Progression: CD4 for monitoring HIV Progression 31st Annual National Conference of Indian Association of Preventive & Social Medicine, 27th – 29th Feb 2004 PGIMER, Chandigarh Dr. Arvinder Singh BD BiosciencesBD Activities: Brazil -COOPEX -MOH -BDB BD ActivitiesSlide3: HIV/AIDS What is CD4/CD8Flow Cytometry Clinical Applications: Flow Cytometry Clinical Applications HIV immunophenotyping (CD4 absolute counts) Cytokine analysis Leukemia and lymphoma diagnosis - minimum residual disease detection Cell cycle and DNA ploidy analysis Stem Cell Analysis HLA – B 27 Typing Residual white blood cell detection Reticulocyte enumeration Flow crossmatching (organ transplantation)What Is Flow Cytometry?: What Is Flow Cytometry? A technology that simultaneously measures multiple characteristics of single cells at a rapid rate Lysed Whole Blood: Lysed Whole Blood Neutrophils Monocytes Lymphocytes Side Scatter Forward Light ScatterTwo-Color Cell Analysis: Two-Color Cell Analysis CD8 PE CD4 FITC 10 0 10 0 10 1 10 2 10 3 10 4 10 1 10 2 10 3 10 4Slide14: Four color analysis CD4 Cy5 CD8 PE CD45 PerCP CD3 FITC CD8 PE CD3 FITC SSC Height CD4 Cy5 Slide15: 42 Million People Living with AIDSIndia – Current Situation HIV: India – Current Situation HIV HIV prevalence rate is 0.7% India has had a sharp increase in the estimated number of HIV infections, from a few thousand in the early 1990s to a working estimate of about 3.8 million children and adults living with HIV/AIDS in 2001 Estimates range from 8 - 15 million HIV infected adults and children living with HIV in 2010 Given India's large population, a mere 0.1 percent increase in the prevalence rate would increase the number of adults living with HIV/AIDS by over half a million peopleIndia – Current Situation HIV: India – Current Situation HIV About 90% of the total reported AIDS cases occur in the sexually active and economically productive 15 to 44 age group Men account for 79% of HIV infections in India The predominant mode of HIV transmission is through heterosexual contact In 2001, the HIV infection rate went above one per cent in six states Maharashtra, Tamil Nadu and Manipur - account for 75% of estimated HIV cases the burden of AIDS cases is beginning to be felt in states affected early - Mumbai and Manipur have recorded 20 to 49 per cent bed occupancy by HIV positive people in certain hospitalsHIV/AIDS Pandemic: HIV/AIDS Pandemic Disease Management Vaccines, Drugs, Education Antiretroviral Therapy (ART) Controls disease progression Increasing drug resistance patterns Expensive (Big Pharma – western countries) Trend to affordable (generic) drugs in developing world CD4 T-Lymphocyte Count Flow cytometry is gold standard Alternate technologies – currently inadequate FacsCount is considered system of choice CD4 count is the single most important objective criteria to guide clinical decision-making on when to use ART HIV Testing paradigm: HIV Testing paradigm Screen Confirm* CD4 Viral load 15-20M 1-2M 4-6M 5-8M Est. annual tests *(includes genotyping)PCR Viral load: PCR Viral load It is a technique that imitates a cell’s ability to replicate DNA by generating multiple copies of specific sequences of DNA through amplification. In clinical diagnostics, a specimen of genetic material can be repeatedly copied by PCR to provide sufficient material to detect the presence or absence of a virus as well as to quantitate its levels in the blood. It’s used for: The development of new drugs To establish a patient’s disease status and the likelihood of disease progression To determine when to start treatment To help the physician decide which drugs to use in a treatment regimen To assess whether a treatment regimen is working CD4 and viral load: CD4 and viral loadHIV/AIDS Pandemic: HIV/AIDS Pandemic Situational Analysis Unmet Needs Affordable drugs/ART Affordable CD4 Flow Cytometry: Flow Cytometry GOLD STANDARD FOR CD4/CD8 Monitoring in HIV/AIDS Patients Conceptually established by BD WorldwideFACSCount - Complete SystemIVD approved ( US FDA ): FACSCount - Complete System IVD approved ( US FDA )FACSCount - Complete System: Premeasured Reagents FACSCount - Complete SystemFACSCount - Complete System: FACSCount Reagents and Controls FACSCount Workstation FACSCount - Complete SystemFACSCount: FACSCount Measuring Absolute CD4, CD8 and CD3 Counts In Three Easy StepsFACSCount - Clinical Need: Safety Minimal Sample Preparation Fixed Samples Absolute Counts Limited Handling of Biohazardous Samples FACSCount - Clinical NeedFACSCount - Clinical Need: Easy to use Absolute Counts Unit Dose Reagents Minimal Hands-On Time No Formal Training Required FACSCount - Clinical NeedFACSCounts in India: 35 Placements 23 Instruments through NACO 12 Instruments in Private Path Laboratories FACSCounts in IndiaReference Range of Lymphocytes Subsets In Healthy Individuals (India): Reference Range of Lymphocytes Subsets In Healthy Individuals (India) MALES CD3+ 617 – 2485 CD19+ 125 – 701 CD4+ 337 – 1090 CD8+ 174 – 1240 Ratio 0.61 – 2.68 HIV Patients-Low CD4 FEMALES CD3+ 898 – 2786 CD19+ 119 – 721 CD4+ 424 – 1050 CD8+ 255 – 1353 Ratio 0.95 – 2.40FACSCount - Analysis: A hard copy results The control Patient sample FACSCount - AnalysisHIV Monitoring: HIV Monitoring Appropriate Technology Flow Cytometry Capacity Building Training, Support, Service BD provides a gold standard system of reliable products and support services for CD4+ T-cell testing at affordable pricing. Reliable Laboratory Systems GMP/GLP (IVDP)/GCP Performance of Competitive Tests: Performance of Competitive TestsPharmas taking action: Pharmas taking action Abbott Laboratories Bristol-Meyers Squibb GlaxoSmithKline Hoffmann La-Roche Ltd Boehringer Ingelheim Merck & Co., Inc Pfizer Cipla Ranbaxy MatrixSlide36: General concensus is that flow cytometry based CD4 count is the most accepted technique to HIV monitoring Quantitative – more cells counted, more reliable result Specific – mAb, identify CD4 T-lymphocytes most precisely QA/QC – internal and external - necessary for accreditation Throughput – manual operation is acceptable as technician costs are not significant and does not limit test demand Existing flow cytometry systems (FACSCalibur or similar) are adequate/accepted for Central Reference facilities – usually at the sites of principal government services Perform broad clinical testing plus Research studies associated with international partners Affordable lower cost platform required at Provincial and Local clinic sites – dedicated clinical test platform Capability to perform CD4 count ( - ideal with broader test menu ) Current FACSCount system should be viewed as maximal technology complexity – sample preparation potential issue – reagent cost limitation Challenge from Partec / Guava like technology or alternative assays Application Comparison: Application ComparisonSlide38: Affordable Testing for HIV/AIDS in the Developing World February 5, 2004 On January 14, 2004, the William J. Clinton Presidential Foundation announced that BD, through BD Biosciences, had entered into an agreement with the Foundation to provide affordable CD4 testing, both reagents and instruments, to the poorest and most severely HIV/AIDS afflicted areas in the developing world. The CD4 immunocytometry test has been used to establish decision points for antiretroviral (ARV) therapy, to monitor efficacy of various treatments, as a prognostic indicator, and as one criterion for initiating prophylaxis for opportunistic infections. The viral load test helps measure how effective ARV therapies are in suppressing the virus and can alert clinicians about the need to adjust dosages or change regimens. BD BiosciencesAccomplishing our Mission – HIV/AIDS: BD Biosciences Accomplishing our Mission – HIV/AIDS BD Biosciences is Helping With the Global AIDS Crisis by: Supporting the Brazilian AIDS prevention program Example program for reducing mortality and new infections in the developing world Our flow instruments and reagents are used as the gold standard to monitor disease progression in HIV-infected individuals Collaborating with the International AIDS Vaccine Initiative (IAVI) Speed the development and distribution of preventive AIDS vaccines Our flow instruments and reagents are used to detect immune response to the vaccineHIV/AIDS : HIV/AIDS 2010 80+ million PLWHA India (15+ million) China (15+ million) Negative growth rates in most of Africa due to AIDS mortality WW economic impact 2002 Brazil Achieves 50% reduction in AIDS deaths over last 10 years Accessible drugs and health care $500,000 million savings on AIDS impact PANDEMIC You do not have the permission to view this presentation. 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