AEGiS-13IAC: Dried blood spot DNA & RNA PCR for infant diagnosis of HIV-1 infection in Thailand.

13th International AIDS Conference

Durban, South Africa - July 9-July 14, 2000

Dried blood spot DNA & RNA PCR for infant diagnosis of HIV-1 infection in Thailand.

Int Conf AIDS 2000 Jul 9-14; 13:(abstract no. MoOrA110)

Young NL, Sangtaweesin V, Chaowanachan T, Chokephaibulkit K, Wasinrapee P, Neeyapun K, Jetsawang B, Teeraratkul A, Mastro TD
N.L. Young, HIV/AIDS Collaboration and US Centers for Disease, Control, DMS6 Building MOPH, Tivanon Road, Nonthaburi 11000, Thailand, Tel.: +66 2 591 5444, Fax: +66 2 580 0696, E-mail: nlyo@cdc.gov

BACKGROUND: It is simpler to collect, store, and transport blood specimens using filter paper collections (filter- paper) rather than venipuncture collections into blood tubes (blood). We compared HIV DNA and RNA PCR results of specimens collected by filter-paper and blood from babies born to HIV-infected women in Thailand.

METHODS: 146 specimens (convenience sample) were collected by both filter-paper and blood from infants and children ages 2 days-52 months (median 4 months) being followed in 2 large Bangkok hospitals. All blood specimens were collected into EDTA blood tubes and were tested using the modified (reduced sample input and annealing temperature) Roche Amplicor HIV-1 DNA PCR test kit version (v.)1.0. For the 25 blood specimens that tested positive and the first 33 blood specimens that tested negative, corresponding filter-paper specimens were also tested using the same kit v.1.0, plus the v.1.5 kit. These same 58 samples also had their blood and filter-paper samples tested using the Organon Teknika NucliSens qualitative RNA PCR test. The 25 blood positives additionally had their filter-paper samples tested using the Roche Amplicor Monitor quantitative RNA PCR test, with the filter-paper copies/mL HCT-and constant factor-corrected. All filter-paper samples were silica-particle extracted for both RNA PCR methods.

RESULTS: All 33 DNA PCR negative blood samples were also negative by DNA and qualitative RNA PCR on matching filter-paper samples. For the 25 filter-paper blood specimens that were DNA PCR positive, matching filter-paper specimens were DNA PCR positive for 20 (80%) specimens using the Amplicor v.1.0, and for 25 (100%) specimens using v.1.5. All 25 filter-paper specimens were positive by qualitative RNA PCR and had detectable RNA by quantitative RNA PCR. The median log difference between the blood and filter-paper HCT-corrected viral loads was 0.27, and the median log difference between the filter-paperHCT-corrected and the filter-paper constant-factor correct viral loads was 0.15.

CONCLUSIONS: Specimens collected onto filter-paper from HIV-exposed infants can be used to detect HIV DNA and RNA, as well as quantify RNA accurately, which would be particularly useful in developing countries.

Keywords: AEGIS, HIV Infections, Polymerase Chain Reaction, RNA, Viral Load, DNA, DNA, Viral, HIV, RNA, Viral, HIV Antibodies, Branched DNA Signal Amplification Assay, Anti-HIV Agents, AIDS Serodiagnosis, HIV Seropositivity, HIV Seroprevalence, Blood Specimen Collection, Filtration, Thailand, Case-Control Studies, Infant, Human, Female, Child, diagnosis, blood, immunology
000709
MoOrA110