Enzyme-linked immunosorbent assay-format microneutralization test for dengue viruses.

Vorndam, Vance; Beltrán, Manuela

doi:10.4269/ajtmh.2002.66.208

Cited by 70 publications

(64 citation statements)

References 16 publications

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“…MNT, as described by Vorndam and Beltran, 18 was performed on paired specimens from study participants whose enrollment anti-DENV IgG ELISA was positive at a 1:100 dilution or specimen pairs from participants with an IgG anti-DENV ELISA titer 1:40 in any specimen. Briefly, 96-well plates were seeded with Vero cells (ATCC) suspended in Enhanced-M199 media at a concentration of 2 + 10 5 cells/mL and incubated at 37 C in 10% CO 2 for 2 days.…”

Section: Methodsmentioning

confidence: 99%

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Incidence of Dengue Virus Infection in School-Aged Children in Puerto Rico: A Prospective Seroepidemiologic Study

Argüello¹,

Tomashek²,

Quiñones³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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Abstract. Dengue is a potentially fatal acute febrile illness caused by the mosquito-borne dengue viruses (DENV-1 to -4). To estimate DENV seroincidence in school-aged children, a 1-year prospective cohort study was conducted in Patillas, Puerto Rico; 10-to 18-year-olds (N = 345) were randomly selected from 13 public schools. At enrollment, 49.8% of the entire cohort had DENV immunoglobulin G (IgG) anti-DENV antibodies, and there were individuals with neutralizing antibodies specific to each of the four DENV. The mean age of participants with incident DENV infection was 13.4 years. The 1-year seroincidence rate was 5.6%, and 61.1% of infections were inapparent. Having IgG anti-DENV at enrollment was associated with seroincidence (risk ratio = 6.8). Acute febrile illnesses during the study period were captured by a fever diary and an enhanced and passive surveillance system in the municipios of Patillas and Guayama. In summary, at enrollment, nearly one-half of the participants had a prior DENV infection, with the highest incidence in the 10-to 11-year-olds, of which most were inapparent infections, and symptomatic infections were considered mild.

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Section: Methodsmentioning

confidence: 99%

“…After incubation, the 96-well plates were fixed in a 1:1 mixture of acetone and methanol for 20 minutes, and the ELISA was performed as previously described. 18 Plates were read on a BioTek plate reader at 405 nm. When the virus control reached an OD value of 1.0-1.3, the plate was read, and the sample was compared with the virus control.…”

Section: Methodsmentioning

confidence: 99%

Incidence of Dengue Virus Infection in School-Aged Children in Puerto Rico: A Prospective Seroepidemiologic Study

Argüello¹,

Tomashek²,

Quiñones³

et al. 2015

The American Society of Tropical Medicine and Hygiene

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“…15 Primary DENV infection was defined by a single reciprocal titer ≥ 40; samples with a reciprocal titer ≥ 40 to more than one DENV type were characterized as having evidence of second or greater DENV infection or having DENV cross-reactive antibodies. Specimens were also tested for evidence of WNV neutralizing antibodies by PRNT 90 .…”

Section: Methodsmentioning

confidence: 99%

Serological Evidence of Infection with Endemic Human Pathogens Among Free-Ranging Old World Monkeys in Puerto Rico

Hemme¹,

Lopez-Ortiz²,

García³

et al. 2016

The American Society of Tropical Medicine and Hygiene

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Abstract. Serum specimens from free-ranging but nonnative patas monkeys (Erythrocebus patas) and rhesus macaques (Macaca mulatta) in southwestern Puerto Rico (PR) were tested for antibodies to infection with dengue viruses (DENVs), West Nile virus (WNV), Leptospira species, and Burkholderia pseudomallei by microneutralization, plaque reduction neutralization, microscopic agglutination, and indirect hemagglutination, respectively. Of 23 animals (21 E. patas and two M. mulatta) tested, all had evidence of prior DENV infection, and of 17 animals tested for WNV, nine (53%) had evidence of prior infection. Of 24 (22 E. patas, two M. mulatta) tested for Leptospira spp., 10 (42%) had evidence of prior exposure, and one patas monkey had antibodies against B. pseudomallei. The acquisition of pathogens endemic among humans in PR by resident nonhuman primates merits further study to define modes of acquisition.

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“…[4][5][6][7][8][9] One assay, a quantitative CmNt for testing antibodies to adenovirus, was considered most applicable for adaptation, offering the advantages of automated, colorimetric results in a microtiter format. 5 CmNt uses the same vital stain used in PRNT, neutral red, as an objective, quantifiable indicator of percent neutralization but obviates the need for an agarose overlay.…”

Section: Introductionmentioning

confidence: 99%

High Throughput Quantitative Colorimetric Microneutralization Assay for the Confirmation and Differentiation of West Nile Virus and St. Louis Encephalitis Virus

Taketa-Graham¹,

Pereira²,

Baylis³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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An automated colorimetric micro-neutralization assay (CmNt) was developed for confirmation and differentiation of West Nile Virus (WNV)-positive human sera as a higher throughput alternative to the standard six-well plaque-reduction neutralization test (PRNT). CmNt was performed in high-capacity 96-well micro-titer plates and required 4–6 days to complete. Inhibition of infection was determined by reduced neutral red-dye retention and conveniently recorded by a colorimetric plate reader. Human sera previously confirmed by PRNT as either negative (N = 52), WNV positive (N = 81), or St. Louis encephalitis virus positive (N = 12) were tested by CmNt; interpreted results were virtually identical to PRNT with a reduced turnaround time and higher throughput. Additionally, a handful of dengue virus positive and negative specimens (four each) were tested by CmNt; interpreted results were identical to PRNT.

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Enzyme-linked immunosorbent assay-format microneutralization test for dengue viruses.

Cited by 70 publications

References 16 publications

Incidence of Dengue Virus Infection in School-Aged Children in Puerto Rico: A Prospective Seroepidemiologic Study

Incidence of Dengue Virus Infection in School-Aged Children in Puerto Rico: A Prospective Seroepidemiologic Study

Serological Evidence of Infection with Endemic Human Pathogens Among Free-Ranging Old World Monkeys in Puerto Rico

High Throughput Quantitative Colorimetric Microneutralization Assay for the Confirmation and Differentiation of West Nile Virus and St. Louis Encephalitis Virus

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