Back to Search
Start Over
Robust and sensitive subtype-generic HIV-1 pol genotyping for use with dried serum spots in epidemiological studies
- Source :
- Journal of Virological Methods. 259:32-38
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- HIV-1 genotyping of larger pol-fragments from dried serum/plasma spots (DSS/DPS) is often hindered by RNA-degradation during transportation at ambient temperature. We evaluated an in-house HIV-1 DSS/DPS-genotyping assay designed in two shorter overlapping fragments covering all resistance mutations in protease and reverse transcriptase. Validation criteria such as specificity, detection limit, accuracy, reproducibility and storage conditions were assessed using reference plasma samples prepared as DPS and clinical DSS from the German molecular HIV-1 surveillance processed under real-life transportation conditions. The specificity was 100% for both samples types, and the experimental DPS detection limit of 1000 copies/ml yielded a 98.7% (3,329/3373) success rate for DSS (including all subtypes) above this detection limit. Accuracy for DPS compared to the gold standard was 99.1% and the reproducibility was 100% for DPS replicates and 99.9% for DSS pairs. Storage of DPS at room temperature was possible for 90 or 30 days and at −20 °C for at least 180 or 90 days at viral loads of 10,000 or 1000 copies/ml, respectively. The HIV-1 pol-genotyping assay presented here is a sensitive, robust and subtype generic tool for a large-scale population-based HIV-1 drug resistance surveillance for the use of DSS/DPS.
- Subjects :
- Serum
0301 basic medicine
Genotyping Techniques
030106 microbiology
Population
HIV Infections
Drug resistance
Biology
Sensitivity and Specificity
Specimen Handling
03 medical and health sciences
0302 clinical medicine
Blood serum
Germany
Virology
Drug Resistance, Viral
Humans
030212 general & internal medicine
Desiccation
education
Genotyping
Detection limit
Reproducibility
education.field_of_study
Chromatography
Reproducibility of Results
Gold standard (test)
pol Gene Products, Human Immunodeficiency Virus
Mutation
HIV-1
Viral load
Subjects
Details
- ISSN :
- 01660934
- Volume :
- 259
- Database :
- OpenAIRE
- Journal :
- Journal of Virological Methods
- Accession number :
- edsair.doi.dedup.....c57d3b50cf0fae9bfc47330d8af267ab