Simple subtyping assay for human immunodeficiency virus type 1 subtypes B, C, CRF01-AE, CRF07-BC, and CRF08-BC

Journal of Clinical Microbiology
Min WeiY Shao

Abstract

After the initial development of a human immunodeficiency virus type 1 (HIV-1) subtype-screening tool by nested multiplex PCR, we further improve it through the redesign of subtype-specific primers based on subtype signature pattern (SSP) analyses and optimization of the PCR conditions. Extracted RNA from plasma samples was used in reverse transcription and the cDNA products were added to the first round PCR, in which universal primers in the gag region were used to detect HIV-1 M group isolates. In the second round of PCR, three pairs of subtype-specific primers, detecting subtypes B, C, and CRF01-AE, were added in one tube. Subtype determination was based on the different size of PCR products on the agarose gel electrophoresis. An additional set of primers detecting only the prevalent recombinant strains CRF07-BC and CRF08-BC was used to discriminate CRF07- and CRF08-BC from pure subtype C. Testing for all kinds of HIV subtype reference strains indicated that this assay was applicable. A panel of 252 HIV-positive samples and 30 HIV-negative samples was further used to evaluate and validate this assay. Compared to the assay of sequence-based phylogenetic analysis, the newly developed assay has an adequate designated subtype se...Continue Reading

References

Sep 1, 1992·AIDS Research and Human Retroviruses·B Korber, G Myers
Jun 10, 2000·Science·B KorberT Bhattacharya
Jun 22, 2000·Molecular and Cellular Probes·L I González-VillaseñorL Yang

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Citations

Feb 9, 2008·AIDS Research and Human Retroviruses·Zhefeng MengYiming Shao
Aug 12, 2016·International Journal of Environmental Research and Public Health·Yansheng YanXun Lin
Dec 24, 2014·The American Journal of Tropical Medicine and Hygiene·Jack Bee ChookKok Keng Tee
Apr 4, 2006·Revista de saúde pública·Henry I Z Requejo
Feb 5, 2010·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Yen-Ju ChenYi-Ming Arthur Chen
Jun 24, 2015·Journal of Acquired Immune Deficiency Syndromes : JAIDS·Min WeiYiming Shao
Apr 25, 2020·International Journal of Molecular Sciences·Wen-Hung WangSheng-Fan Wang
Apr 14, 2016·AIDS Research and Human Retroviruses·Danying ChenXia Feng

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