Multiplex Analysis of Heterophil Antibodies in Patients With Indeterminate HIV Immunoassay Results

Willman, Joseph H.; Hill, Harry R.; Martins, Thomas B.; Jaskowski, Troy D.; Ashwood, Edward R.; Litwin, Christine M.

doi:10.1309/3f13-739a-na7f-nv3x

Cited by 38 publications

(18 citation statements)

References 19 publications

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“…The Western blot has traditionally been the gold standard against which HIV assays are compared. It should be noted, however, that false-positive Western blots have been reported (7,17,32,40) and that when comparing tests to a gold standard, the best a test can do is match the Western blot results. If the test is actually better or if the Western blot is incorrect in some instances, a more accurate test would look worse.…”

Section: Resultsmentioning

confidence: 99%

“…The causes of indeterminate patterns on Western blot include technical artifact, laboratory error, irrelevant cross-reactions, nonspecific binding, infection with a related retrovirus such as HIV-2, the presence of an antigenic variant of HIV-1, early HIV-1 infection, or late-stage disease (3,6,7,17,32,40). Antiretroviral therapy reduces viral load and has been reported to reduce HIV-specific antibody (11,16,22,23,28).…”

Section: Resultsmentioning

confidence: 99%

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Alternative Algorithms for Human Immunodeficiency Virus Infection Diagnosis Using Tests That Are Licensed in the United States

et al. 2008

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Serodiagnosis of human immunodeficiency virus (HIV) infection in the UnitedStates has traditionally relied on a sequential two-test algorithm: an initial screen with an enzyme immunoassay (EIA) and reflex testing of EIA-reactive specimens with a more specific supplemental test such as Western blotting or immunofluorescence. The supplemental tests are tedious, subjective, and expensive. In addition, there have been major improvements in the performance and accuracy of the EIA tests as well as the introduction of rapid serologic tests (RT) and HIV nucleic acid amplification tests (NAAT). Related to these improvements is the possibility that alternative algorithms using combinations of currently approved HIV tests may function as well as if not better than the current algorithm, with more flexibility, improved accuracy, and lower cost. To this end, we evaluated the performance of 12 currently licensed tests and 1 in-house HIV test (6 EIA, 4 RT, and 3 NAAT) on panels of plasma samples from HIV-infected (n ‫؍‬ 621 HIV type 1 [HIV-1] and 34 HIV-2) and uninfected (n ‫؍‬ 513) people and of sequential specimens from people early in seroconversion (183 specimens from 15 patients). Test combinations were analyzed in two dual-test (sensitivity-optimized and specificity-optimized) algorithms and in a three-test (tie-breaking) algorithm, and performance was compared to the conventional algorithm. The results indicate that alternative algorithm strategies with currently licensed tests compare favorably with the conventional algorithm in detecting and confirming established HIV infection. Furthermore, there was a lower frequency of discordant or indeterminate results that require follow-up testing, and there was improved detection of early infection.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Alternative Algorithms for Human Immunodeficiency Virus Infection Diagnosis Using Tests That Are Licensed in the United States

et al. 2008

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“…The SPHL panel had only 44 IgM negatives of the 583 tested by using the CDC protocol; however, 14 of the 44 CDC IgM protocol negative samples were positive by the Focus IgM assay. As with the IgG samples in the SPHL serum panel, the apparent lack of IgM specificity by the Focus assay may be due to lack of sensitivity of the CDC protocol performed at 16 Heterophile antibodies in human sera react with proteins from other mammalian species; examples include human antimouse immunoglobulin, human anti-horse immunoglobulin, and human anti-bovine albumin (5,11,25). The presence of heterophile antibodies is known to interfere in numerous serologic assays, in particular IgM assays (5,11).…”

Section: Discussionmentioning

confidence: 99%

Performance of Immunoglobulin G (IgG) and IgM Enzyme-Linked Immunosorbent Assays Using a West Nile Virus Recombinant Antigen (preM/E) for Detection of West Nile Virus- and Other Flavivirus-Specific Antibodies

Hogrefe¹,

Moore²,

Lapé-Nixon³

et al. 2004

J Clin Microbiol

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Focus Technologies developed an indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay(ELISA) and a mu-capture IgM ELISA for the detection of West Nile virus (WNV)-specific antibodies based on a WNV preM/E protein recombinant antigen. Normal and disease state serum panels were used to assess the performance characteristics of the two WNV ELISA kits. Totals of 807 and 1,423 sera were used to assess the IgG ELISA and IgM ELISA kits, respectively. The Focus Technologies IgG ELISA had a sensitivity of 97.6% and a specificity of 92.1% (excluding non-WNV flavivirus sera). The comparative method for WNV IgG may lack sensitivity in detecting IgG in early WNV infection, so the specificity of the Focus IgG ELISA may be higher than 92.1%. When sera from patients either infected with or vaccinated against other flaviviruses were tested on the WNV IgG assay, 35% of the sera reacted as positive for WNV IgG. Yellow fever and Japanese encephalitis vaccinees were less reactive in the IgG ELISA than St. Louis and dengue fever patients. The Focus Technologies IgM ELISA had a sensitivity and a specificity of 99.3% (excluding the non-WNV flavivirus sera). The overall cross-reactivity for the IgM ELISA to flavivirus sera was 12%, with 31% of St. Louis encephalitis patients found to be WNV IgM positive and no yellow fever vaccinees found to be WNV IgM positive. In a selected population of 706 sera, 15 false-positive WNV IgM sera were identified. The use of a background subtraction method for the IgM ELISA eliminated all 15 false-positive results, giving a specificity of 100% for the Focus IgM ELISA.Since the initial outbreak of West Nile virus (WNV) in New York in 1999, WNV has spread rapidly across the entire continental United States in only 4 years (3,7,13,(16)(17)(18). WNV serology, in particular the detection of WNV immunoglobulin M (IgM) in both serum and cerebrospinal fluid, has become the primary tool for diagnosing human WNV infection. The detection of WNV IgM in serum represents a probable WNV infection, whereas the detection WNV IgM in cerebrospinal fluid is considered diagnostic of central nervous system involvement by WNV (13,15,24). Due to very low viremia at the time of clinical onset, nucleic acid detection methods and WNV culture are not useful diagnostic tools (10, 13). Only 20% of WNV-infected individuals are symptomatic; the majority of symptomatic patients present with a self-limited viral syndrome of fever, headache, malaise, and rash. Fewer than 1% of infected individuals progress to serious clinical disease, typically manifesting as either meningitis or encephalitis (19,22).WNV is a member of the Flaviviridae family and is in the Japanese encephalitis serocomplex that includes Japanese encephalitis (JE) virus and St. Louis encephalitis (SLE) virus. Other closely related flaviviruses include yellow fever (YF) virus and dengue virus types 1 to 4. The flavivirus antibody response is predominantly generated against the highly immunogenic envelope protein that contains both flavivirus crossreactive epitop...

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“…Ags were coupled to the microspheres using a modification of the protocol from Luminex Corp. Aliquots of 5 million SeroMAP Microspheres (Luminex Corp., Austin, TX) with different spectral addresses (17,18,19,21,22,44,50,51,59, 64, and 69) were centrifuged at 8,000 rpm for 2 min, washed with 100 l of distilled water, resuspended in 80 l of activation buffer (0.1 M NaH 2 PO 4 , pH 6.2), and then activated with 10 l of a 50-mg/ml solution of 1-ethyl 3-(3-dimethylaminopropyl) carbodiimide hydrochloride and 10 l of a 50-mg/ml solution of sulfo-N-hyroxysulfosuccinimide. The microspheres were mixed gently and incubated at room temperature (RT) in the dark for 20 min.…”

Section: Methodsmentioning

confidence: 99%

“…The technique has been used to measure Ab levels to protein Ags of several microbes (12,14,17) but not for more complex parasites. SAT was reported to give results comparable to those of ELISA for quantification of Abs against tetanus, diphtheria, Haemophilus influenzae type B (13), pneumococcal capsular polysaccharides (9,14), and Neisseria meningitidis (8).…”

mentioning

confidence: 99%

Multiplex Assay for Simultaneous Measurement of Antibodies to Multiple Plasmodium falciparum Antigens

Fouda

Leke

Long

et al. 2006

Clin Vaccine Immunol

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Antibodies to Plasmodium falciparum are classically measured using the enzyme-linked immunosorbent assay (ELISA). Although highly sensitive, this technique is labor-intensive when large numbers of samples must be screened against multiple antigens. The suspension array technology (SAT) might be an alterative to ELISA, as it allows measurement of antibodies against multiple antigens simultaneously with a small volume of sample. This study sought to adapt the new SAT multiplex system for measuring antibodies against nine malarial vaccine candidate antigens, including recombinant proteins from two variants of merozoite surface protein 1, two variants of apical merozoite antigen 1, erythrocyte binding antigen 175, merozoite surface protein 3, and peptides from the circumsporozoite protein, ring erythrocyte surface antigen, and liver-stage antigen 1. Various concentrations of the antigens were coupled to microspheres with different spectral addresses, and plasma samples from Cameroonian adults were screened by SAT in mono-and multiplex formats and by ELISA. Optimal amounts of protein required to perform the SAT assay were 10-to 100-fold less than that needed for ELISA. Excellent agreement was found between the single and multiplex formats (R > 0.96), even when two variants of the same antigen were used. The multiplex assay was rapid, reproducible, required less than 1 l of plasma, and had a good correlation with ELISA. Thus, SAT provides an important new tool for studying the immune response to malaria rapidly and efficiently in large populations, even when the amount of plasma available is limited, e.g., in studies of neonates or finger-prick blood.

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Multiplex Analysis of Heterophil Antibodies in Patients With Indeterminate HIV Immunoassay Results

Cited by 38 publications

References 19 publications

Alternative Algorithms for Human Immunodeficiency Virus Infection Diagnosis Using Tests That Are Licensed in the United States

Alternative Algorithms for Human Immunodeficiency Virus Infection Diagnosis Using Tests That Are Licensed in the United States

Performance of Immunoglobulin G (IgG) and IgM Enzyme-Linked Immunosorbent Assays Using a West Nile Virus Recombinant Antigen (preM/E) for Detection of West Nile Virus- and Other Flavivirus-Specific Antibodies

Multiplex Assay for Simultaneous Measurement of Antibodies to Multiple Plasmodium falciparum Antigens

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