Feasibility of pooling sera for HIV-1 viral RNA to diagnose acute primary HIV-1 infection and estimate HIV incidence

Quinn, Thomas C.; Brookmeyer, Ron; Kline, Richard L.; Shepherd, Mary; Paranjape, Ramesh; Mehendale, Sanjay; Gadkari, Deepak A.; Bollinger, Robert C.

doi:10.1097/00002030-200012010-00015

Cited by 118 publications

(100 citation statements)

References 30 publications

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“…Detection methods based on pooled nucleic acid could provide a simple solution. The pooling of sera or plasma specimens has long been used to improve the accuracy, costeffectiveness, and throughput of screening for many infections (e.g., HIV-1, hepatitis B virus, hepatitis C virus, and West Nile virus) in low-prevalence settings, such as blood banks (4,26). We recently reported methods for pooled analysis of dried blood spots for HIV-1 nucleic acids (1,18).…”

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confidence: 99%

PCR-Based Pooling of Dried Blood Spots for Detection of Malaria Parasites: Optimization and Application to a Cohort of Ugandan Children

Hsiang¹,

Lin²,

Dokomajilar

et al. 2010

J Clin Microbiol

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Sensitive, high-throughput methods to detect malaria parasites in low-transmission settings are needed. PCR-based pooling strategies may offer a solution. We first used laboratory-prepared samples to compare 2 DNA extraction and 4 PCR detection methods across a range of pool sizes and parasite densities. Pooled Chelex extraction of DNA, followed by nested PCR of cytochrome b, was the optimal strategy, allowing reliable detection of a single low-parasitemic sample (100 parasites/l) in pool sizes up to 50. This PCR-based pooling strategy was then compared with microscopy using 891 dried blood spots from a cohort of 77 Ugandan children followed for 2 years in an urban setting of low endemicity. Among 419 febrile episodes, 35 cases of malaria were detected using the PCR-based pooling strategy and 40 cases using microscopy. All five cases of malaria not detected by PCR were from samples stored for >2 years with parasitemia of <6,000/l, highlighting the issue of possible DNA degradation with long-term storage of samples. Among 472 samples collected from asymptomatic children as part of routine surveillance, 15 (3.2%) were positive by PCR-based pooling compared to 4 (0.8%) by microscopy (P ‫؍‬ 0.01). Thus, this PCR-based pooling strategy for detection of malaria parasites using dried blood spots offers a sensitive and efficient approach for malaria surveillance in low-transmission settings, enabling improved detection of asymptomatic submicroscopic infections and dramatic savings in labor and costs.Due to large-scale implementation of effective control measures, many countries where malaria is endemic are experiencing dramatic declines in disease burden. With this success has come a shift in the end goal from control to elimination (9, 10). However, when the goal is elimination, accurate detection of persons infected with malaria parasites becomes essential (11). Standard surveillance systems depend on diagnosis by microscopy, a method that is technically challenging, labor-intensive, and often inaccurate in operational settings. More recently available rapid diagnostic tests (RDTs) provide convenience and ease of use, but they have limitations in specificity, sensitivity, species identification, and cost (22).PCR-based methods for malaria parasite detection are relatively simple and provide improved sensitivity compared to microscopy and RDTs, especially in settings where infections have low parasitemia or contain mixed species (22). PCR can also be performed on dried blood spots, which are convenient for collection, storage, and transport. Nonetheless, PCR has a long turnaround time, making it an impractical tool for clinical care of individual patients. One exception would be real-time PCR, which has a short turnaround time but carries with it cost and capacity constraints that make it unavailable for rapid diagnosis in most settings. However, there is a potential role for PCR in situations where large numbers of samples are being screened: a high-throughput system could allow accurate, rapid, and cost-effective ass...

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PCR-Based Pooling of Dried Blood Spots for Detection of Malaria Parasites: Optimization and Application to a Cohort of Ugandan Children

Hsiang¹,

Lin²,

Dokomajilar

et al. 2010

J Clin Microbiol

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“…659,2003). Although some efforts have been directed at detecting p24 antigen or HIV-1 RNA in the absence of HIV antibodies (1,4,10,12,13), other approaches are based on qualitative and quantitative differences in the evolution of HIV antibodies following seroconversion (2, 9, 11; Jenner et al, 10th Conf. Retrovir.…”

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Performance Characteristics of the Immunoglobulin G-Capture BED-Enzyme Immunoassay, an Assay To Detect Recent Human Immunodeficiency Virus Type 1 Seroconversion

et al. 2004

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Recently, we developed an immunoglobulin G (IgG)-capture BED-enzyme immunoassay (BED-CEIA) to identify recent human immunodeficiency virus (HIV) type 1 (HIV-1) seroconversion for use in incidence estimates. We have established an algorithm for its use; developed quality control reagents to monitor the assay; and evaluated its performance for interrun, intrarun, and operator variability. Analysis of 144 individual plates, which involved multiple plate lots and several operators over more than a year, indicated that the coefficients of variation (CVs) were between 10 and 15% for raw optical density (OD) values in the dynamic range between 0.5 and 2.0 OD units; the CVs decreased to 5 to 10% when the OD was normalized (OD-n; OD-n ‫؍‬ specimen OD/calibrator OD). The intrarun CVs were generally in the range of 5 to 10% for specimens with ODs <0.5 and less than 5% for specimens with ODs >0.5. The level of concordance between multiple plate lots (n ‫؍‬ 6) and multiple operators (n ‫؍‬ 7) was quite high (R 2 > 0. 9). Comparison of the results of the initial and the confirmatory tests with specimens with OD-n values

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“…14, S2). Pooling strategies for HIV detection in clinical testing have recently been piloted in Swiss (15), Indian (16), and US (14) clinical-HIV-testing populations. The results suggest that HIV RNA testing -if based on pooling of antibody-negative specimens -can efficiently identify acute HIV infections in clinical testing.…”

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Acute HIV revisited: new opportunities for treatment and prevention

Pilcher¹,

Eron²,

Galvin³

et al. 2004

J. Clin. Invest.

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Inability to recognize incident infection has traditionally limited both scientific and public health approaches to HIV disease. Recently, some laboratories have begun adding HIV nucleic acid amplification testing to HIV diagnostic testing algorithms so that acute (antibody-negative) HIV infections can be routinely detected within the first 1–3 weeks of exposure. In this review article, we will highlight critical opportunities for HIV treatment and prevention that are presented by these diagnostic strategies

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Feasibility of pooling sera for HIV-1 viral RNA to diagnose acute primary HIV-1 infection and estimate HIV incidence

Cited by 118 publications

References 30 publications

PCR-Based Pooling of Dried Blood Spots for Detection of Malaria Parasites: Optimization and Application to a Cohort of Ugandan Children

PCR-Based Pooling of Dried Blood Spots for Detection of Malaria Parasites: Optimization and Application to a Cohort of Ugandan Children

Performance Characteristics of the Immunoglobulin G-Capture BED-Enzyme Immunoassay, an Assay To Detect Recent Human Immunodeficiency Virus Type 1 Seroconversion

Acute HIV revisited: new opportunities for treatment and prevention

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