Detection of HIV-1 in alternative specimen types using the APTIMA ® HIV-1 RNA Qualitative Assay

• Published in: Journal of virological methods Vol. 159; no. 1; pp. 10 - 14
• Main Authors: Nugent, C. Thomas, Dockter, J., Bernardin, F., Hecht, R., Smith, D., Delwart, E., Pilcher, C., Richman, D., Busch, M., Giachetti, C.
• Format: Journal Article
• Language: English
• Published: Kidlington Elsevier B.V 01.07.2009; Elsevier
• Subjects: Biological and medical sciences; Blood Stains; Dried blood spots; Fundamental and applied biological sciences. Psychology; HIV Infections - diagnosis; HIV Infections - genetics; HIV Infections - virology; HIV-1; HIV-1 - genetics; HIV-1 - isolation & purification; Humans; Leukocytes, Mononuclear - virology; Microbiology; Reagent Kits, Diagnostic; RNA detection; RNA, Viral - genetics; RNA, Viral - isolation & purification; Saliva - virology; Semen - virology; Sensitivity and Specificity; Techniques used in virology; Virology; HIV-1; RNA detection; Dried blood spots; Microbiology; HIV-1 virus; Retroviridae; Method; Lentivirus; Blood; Virology; Virus; Human immunodeficiency virus; Detection
• ISSN: 0166-0934; 1879-0984
• DOI: 10.1016/j.jviromet.2009.02.015

Peripheral blood mononuclear cells (PBMCs), saliva, seminal plasma, and dried blood spots were evaluated as specimen types for the APTIMA HIV-1 RNA Qualitative Assay (APTIMA HIV-1 Assay), which employs a target capture step to recover HIV-1-specific sequences from complex specimen types. Analytical sensitivity studies were carried out using samples that were either diluted or eluted with a buffered detergent and spiked with different concentrations of HIV-1 ranging from 1 to 10,000 copies/mL. PBMC samples spiked with HIV-1 had comparable analytical sensitivity to HIV-1 spiked plasma with a 95% limit of detection of 13.1 and 17.2 copies/mL, respectively. Analytical sensitivity in seminal plasma specimens diluted 1:5 and saliva diluted 1:2 was comparable to HIV-1 spiked dilution buffer alone. Whole blood and dried blood spot specimens spiked with HIV-1 had equivalent reactivity at 250 copies/spot (5000 copies/mL). However, the 95% limit of detection values were significantly different (293.7 copies/mL for whole blood and 2384 copies/mL for dried blood spot specimens). No significant effect on analytical sensitivity was observed when one HIV-1 positive dried blood spot punch was pooled with up to 9 HIV-1 negative dried blood spot punches. Together, these studies demonstrate that the APTIMA HIV-1 RNA Qualitative Assay can be used to process a diverse array of specimen types with minimal impact on analytical sensitivity for most specimen types.