Screening for Human Immunodeficiency Virus Infection by Use of a Fourth-Generation Antigen/Antibody Assay and Dried Blood Spots: In-Depth Analysis of Sensitivity and Performance Assessment in a Cross-Sectional Study

Abstract

We evaluated the performance of a fourth-generation antigen/antibody (Ag/Ab) assay for detecting HIV-1 infection on dried blood spots (DBS) both in a conventional laboratory environment and in an epidemiological survey corresponding to a real-life situation. Although a 2-log loss of sensitivity compared to that with plasma was observed when using DBS in an analytical analysis, the median delay of positivity between DBS and crude serum during the early phase postacute infection was 7 days. The performance of the fourth-generation assay on DBS was approximately similar to that of a third-generation (antibody only) assay using crude serum samples. Among 2,646 participants of a cross-sectional study in a population of men having sex with men, 428 DBS were found reactive, but negative results were obtained from 5 DBS collected from individuals who self-reported a positive HIV status, confirmed by detection of antiretroviral (ARV) drugs in their DBS. The data generated allowed us to estimate a sensitivity of 98.8% of the fourth-generation assay/DBS strategy in a high-risk population, even including a broad majority of individuals on ARV treatment among those HIV positive. Our study brings additional proofs that DBS testing using a fourth-generation immunoassay is a reliable strategy able to provide alternative approaches for both individual HIV testing and surveillance of various populations.

Reliability of dried blood spot (DBS) cards in antibody measurement: A systematic review

“…Thirty-two (89%) reported the methodology used to elute the DBS samples [14,15,[17][18][19][20][21][22][23][24][25][26][27][28][29][30][31][32][35][36][37][39][40][41][42][44][45][46][47][48][49][50][51][52]. The shortest incubation period of the DBS in elution buffer (50 μl 0.05% PBS/Tween-20) was 30 minutes [42] and the longest incubation period of the DBS in elution buffer (700 μl and 300 μl PBS/0.05% BSA) was 18-24 hours [17]) ( Table 2).…”

“…Overall, DBS and serum samples from 16,255 individuals were included: 13,742 (84.5%) adults, 560 (3.4%) 5-to 17-year old's and 2,082 (12.8%) less than five years old. Two studies reported antibodies against hepatitis A [14,15], nine hepatitis B [16][17][18][26][27][28]46,48,51], ten hepatitis C [19][20][21][22][26][27][28]37,46,48], eight HIV [23][24][25][26][27][28]46,50], three human papillomavirus (HPV) [29,30,37], three measles [31,32,42], three rubella [33,34,42], two syphilis [40,46], two H. pylori [35,47] and two malaria [38,49]. Twelve papers reported on Chagas disease [35], Epstein-Barr virus [36], HPV, H. pylori, hepatitis C and polyomavirus [37], Strong...…”

“…Twelve papers reported on Chagas disease [35], Epstein-Barr virus [36], HPV, H. pylori, hepatitis C and polyomavirus [37], Strongyloidiasis [39], pertussis [41], hepatitis E [56], Vibrio cholera [2], measles, mumps and rubella [42], tuberculosis and cytomegalovirus [43], toxoplasmosis [44], trypanosoma [45], Covid-19 [52], respectively. Twelve studies were conducted in Europe [14,18,21,27,29,30,35,[41][42][43]50,52], four in North America [19,20,32,36], eleven in South America [15][16][17]22,24,34,38,39,48,49,51], seven in Africa [23,25,26,31,40,44,45] and five in Asia [28,33,…”

“…Thirty of the studies used enzyme-linked immunosorbent assays (ELISA) [14][15][16][17]19,[21][22][23][24][25][26][29][30][31][32]34,36,38,39,[41][42][43][44][45][46][47][49][50][51][52] with the one study using the Luminex 100 [37] or a combination of Treponema pallidum particle agglutination assay (TPPA), Treponema pallidum hemagglutination assay (TPHA) and ELISA [40]. Two studies used the architect system [27,35] as a detection method and two studies used chemiluminescence immunoassay (CIA) either alone [18] or in combination with ELISA [20].…”

Validation of Dried Blood Spot Sample Modifications to Two Commercially Available Covid-19 Igg Antibody Immunoassays

“…The US Department of Transportation and US Postal Service consider DBS to be nonregulated and exempt materials for shipping if they are properly packaged [ 15 ], making it easy to return DBS specimens from a clinic or home directly to a high complexity clinical laboratory for testing. Prior studies have successfully used DBS to detect antibodies against Epstein–Barr [ 16 ], Rabies [ 17 ], Hepatitis B and C [ 18–20 ], HIV [ 18 , 21 , 22 ], Herpes Simplex [ 23 ] and Ebola [ 24 ] viruses.…”