Development of a cost-effective assay for genotyping of HIV-1 non-B subtype for drug resistance.
Highly Active Antiretroviral Therapy (HAART) is the most effective way to control HIV-1 replication in infected patients. Prior to the start of therapy, genotyping of HIV-1 for mutations that confer resistance to potential drug candidates is crucial for it allows formulating an effective regimen. Ineffective drugs are excluded and potentially effective ones are included. A number of diagnostic kits are commercially available for this purpose but are tailored for HIV-1 subtype-B, a strain chiefly found in AIDS patients of Europe and America. However, AIDS patients of South-East Asia including Thailand are predominant infected with HIV-1 subtype non-B. In this study, an inexpensive assay was developed that genotypes HIV-1 non-B for drug resistance and tested it on 99 Thai AIDS patients. Results showed that 98 were infected with HIV-1 subtype non-B (or CRF01_AE) and one with subtype-B. Within the HIV-1 polymerase (pol), reverse transcriptase (RT) gene, the assay identified 18 codon mutations associated with resistance to Nucleoside/Nucleotide Reverse Transcriptase Inhibitors (NRTIs) and 17 Non-Nucleoside Reverse Transcriptase Inhibitors (NNRTIs). Employing a commercially available kit, parallel genotyping of patient samples confirmed results providing validation of the assay. This method approximately costs 100 US dollars compared to $300 for a commercially available test. In Thailand, the burden of cost for treating HIV-infections is high not only for the average citizen but the country's health care systems. Therefore the low cost and yet effective genotyping test for HIV-1 subtype non-B is a practical and viable solution to expensive genotyping platforms.
Department of Transfusion Medicine, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand; Innovation Center for Research and Development of Medical Diagnostic Technology Project, Department of Transfusion Medicine, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand. Electronic address: palanee@gmail.com.
Palanee
P
Ammaranond
Innovation Center for Research and Development of Medical Diagnostic Technology Project, Department of Transfusion Medicine, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, Thailand.
Sayompoo
S
Sanguansittianant
National Engineering Research Center for Miniaturized Detection System, Northwest University, Xi'an, China.
Paul A
PA
Raju
St Vincent's Hospital Sydney Limited, Sydney, Australia.
Philip
P
Cunningham
Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand.
Navin
N
Horthongkham
D024882:Drug Resistance, Viral; D005260:Female; D060005:Genotyping Techniques / Q000191:economics / Q000379:methods*; D015658:HIV Infections / Q000821:virology*; D054303:HIV Reverse Transcriptase / Q000235:genetics; D015497:HIV-1 / Q000187:drug effects* / Q000235:genetics* / Q000302:isolation & purification; D017048:Health Care Costs; D006801:Humans; D008297:Male; D008826:Microbial Sensitivity Tests / Q000191:economics / Q000379:methods; D020125:Mutation, Missense; D013785:Thailand; D054302:pol Gene Products, Human Immunodeficiency Virus / Q000235:genetics
D054302:pol Gene Products, Human Immunodeficiency Virus; C514824:reverse transcriptase, Human immunodeficiency virus 1; D054303:HIV Reverse Transcriptase
Drug resistance; Genotype; HIV-1