[Detection of proviral gene of human immunodeficiency virus type 1 from dried blood spots].
A method for detection of proviral human immunodeficiency virus type 1 DNA fragment encoded pol gene in dried blood spots on filter paper by nested primers polymerase chain reaction (nPCR) has been developed. The filter paper was pre-saturated and dried by EDTA proteineas K. About 50 microliters whole blood from HIV1 infected individuals was collected and onto the piece of filter paper, and the paper was air dried stored sealed in a plastic bag at 4 degrees C or room temperature for 1 to 64 weeks. The PCR with outer primers was performed in 0.5% ml tube with the piece of filter paper. Following the PCR with the inner primers was finished. The results showed that template DNA could detected in piece of filter papers which stored at 4 degrees C for 40 weeks as well as stored at room temperature for 24 weeks. The results could directly determined by ethidium bromide staining of the PCR products when it compared with the PCR products of standard positive control and DNA molecular weight marker in electrophoresis analysis on agarose gel. The method is rapid, specific, and sensitive and has the ability to detect as little as 10 proviral template DNA. The collected samples could mail from basic laboratory to center laboratory and the method could be applied in confirming and screening of HIV1 infection.
Wang
B
B
Laboratory of Molecular Virology Medical College of Qingdao University, Qingdao 266021.
Lu
X
X
Lu
Y
Y
D001801:Blood Stains; D004279:DNA, Viral / Q000032:analysis*; D005814:Genes, Viral / Q000235:genetics*; D015658:HIV Infections / Q000175:diagnosis / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D009000:Monocytes / Q000821:virology*; D016133:Polymerase Chain Reaction
D004279:DNA, Viral