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Contd.
Sample Collection & Workflow:
• Blood: Plasma Viral load and CD4/CD8 count • Semen Aliquot 1, - 2 – 4 ml
→ Sperm preparation through density gradient centrifugation, 2 washes and subsequent swim up.
→ Prepared sperm cryogennically preserved
• Semen Aliquot 2,
→ 1ml of semen to obtain seminal plasma (500uL) by
centrifugation and cell pellet.
Ref. Duru Shah et.al.
Contd. Duru Shah
Contd.
→ Processed by RNA extraction, DNA extraction, subsequent Real Time/Nested-PCR to detect virus.
• Seminalplasmaaswellasspermassociatedvirus assessed using HIV-1-specific PCR (for multiple genomic regions).
• AllprocessingdoneatMetropolis,adiagnosticchain of laboratories (CAP accredited) using appropriate biosafety conditions.
Ref. Duru Shah et.al. Contd. Duru Shah
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PRECONGRESS COURSE 12 I VIENNA, AUSTRIA – 23 JUNE 2019 65
