An effective tool for identifying HIV-1 subtypes B, C, CRF01_AE, their recombinant forms, and dual infections in Southeast Asia by the multi-region subtype specific PCR (MSSP) assay

• Main Authors: Supachai Sakkhachornphop, Gustavo H. Kijak, Chris Beyrer, Myat Htoo Razak, Eric Sanders-Buell, Jaroon Jittiwutikarn, Vinai Suriyanon, Merlin L. Robb, Jerome H. Kim, David D. Celentano, Francine E. McCutchan, Sodsai Tovanabutra
• Format: Journal
• Published: 2018
• Subjects: Immunology and Microbiology
• Online Access: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=84924691983&origin=inward; http://cmuir.cmu.ac.th/jspui/handle/6653943832/54575
• Institution: Chiang Mai University

© 2015 Elsevier B.V. The RV144 Thai vaccine trial has been the only vaccine study to show efficacy in preventing HIV infection. Ongoing molecular surveillance of HIV-1 in Southeast Asia is vital for vaccine development and evaluation. In this study a novel tool, the multi-region subtype specific PCR (MSSP) assay, that was able to identify subtypes B, C, CRF01_AE for Thailand, other Southeast Asian countries, India and China is described. The MSSP assay is based on a nested PCR strategy and amplifies eight short regions distributed along the HIV-1 genome using subtype-specific primers. A panel of 41 clinical DNA samples obtained primarily from opiate users in northern Thailand was used to test the assay performance. The MSSP assay provided 73-100% sensitivity and 100% specificity for the three subtypes in each genome region. The assay was then field-tested on 337 sera from HIV infected northern Thai drug users collected between 1999 and 2002. Subtype distribution was CRF01_AE 77.4% (n = 261), subtype B 3.3% (n = 11), CRF01_AE/B recombinant 12.2% (n = 41), CRF01_AE/C recombinant 0.6% (n = 2), and non-typeable 6.5% (n = 22). The MSSP assay is a simple, cost-effective, and accurate genotyping tool for laboratory settings with limited resources and is sensitive enough to capture the recombinant genomes and dual infections.