Detecting Artificial Anti-Dengue IgM Immune Complexes Using an Enzyme-Linked Immunosorbent Assay

Kuno, Goro; Gómez, Isabel; Gubler, D. J.

doi:10.4269/ajtmh.1987.36.153

Cited by 151 publications

(93 citation statements)

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“…The MAC-ELISA was used as reference and performed, as described earlier. 6 The optical density (OD) values throughout the text represent the means of OD values of two wells per specimen after the mean OD value of the negative control specimen in each plate was subtracted. The standard HI test adapted for microtiter plate was performed, according to the method of Clarke and Casals.…”

Section: Methodsmentioning

confidence: 99%

“…Confirmation of infection was by virus isolation, by the standard hemagglutination inhibition (HI) test on paired serum specimens, and by detecting dengue-specific IgM antibody with the IgM antigen capture ELISA (MAC-ELISA). 6 Ten cerebrospinal fluid (CSF) specimens from serologically confirmed Japanese encephalitis patients from the Philippines and nine serum specimens from serologically and/or virologically confirmed cases of yellow fever in Brazil and Nigeria were also used. For yellow fever infections, only convalescent (Ն 8 days after onset) serum specimens demonstrating a Ն 8-fold higher IgM titer to YF than to DEN virus were used.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Evaluation of an IgM immunoblot kit for dengue diagnosis.

Kuno¹,

Cropp²,

Wong-Lee³

et al. 1998

The American Journal of Tropical Medicine and Hygiene

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Abstract. A commercial IgM immunoblot kit was evaluated for dengue diagnosis with a panel of serum specimens collected from patients in a dengue endemic area. The kit is not recommended for use in its present form because of its undesirable rate of false-positive results. However, by substituting internal controls with the reference positive and negative controls that are more representative of those seen in endemic areas and by modifying the positive and negative scoring criteria, sensitivity and specificity of 80.3% and 94.5%, respectively, were obtained. These results are comparable with those obtained with the IgM ELISA on specimens, most of which were obtained from outpatient health care facilities. With further technical modifications, inclusion of a visual guide to ensure scoring standardization, and a more complete elaboration of the limitations of the test, wide application of the kit in diagnostic laboratories should be possible.Dengue fever is the most important arboviral disease in the world in terms of morbidity and mortality. 1 Incidence of the severe forms of dengue, dengue hemorrhagic fever and dengue shock syndrome (DSS), which emerged in Southeast Asia in the 1950s and 1960s, has increased dramatically in the Americas. Since the first major DHF outbreak in Cuba in 1981, 23 countries have reported confirmed cases of DHF, and outbreaks have occurred in Venezuela, Brazil, Colombia, and French Guiana. 2,3 Although many diagnostic techniques are available, the major problems of dengue diagnosis in many countries in the tropics are high cost and lack of reagents and equipment. Since reagents for most arboviral diseases, including dengue, have not been commercially available until recently, diagnostic services have been limited to a very small number of well-established and equipped laboratories. This is one of the major reasons for poor surveillance, under-reporting of cases, and the long time intervals between the onset of epidemic transmission and the mobilization of mosquito control. For physicians administering medical care, a major problem is the long period between submission of specimens and receipt of results. The recent proliferation of commercial dengue diagnostic kits is highly encouraging in this regard. 4 In particular, the kits that are simple to use are important for field applications in smaller clinics, hospitals, and other health care facilities. Recently, an immunoblot kit that satisfies some of those requirements was evaluated and judged to have excellent qualities in a multicenter study. 5 We have evaluated the same kit but obtained different results. Here, we report those data, discuss the problems of kit evaluation for dengue IgM detection, and present thoughts on how diagnostic kits should be evaluated. . MATERIALS AND METHODSClinical specimens. A panel of 233 serum specimens (96 paired specimens and 137 single specimens) were collected from dengue patients in Puerto Rico. Confirmation of infection was by virus isolation, by the standard hemagglutination inhibition (HI) test o...

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Evaluation of an IgM immunoblot kit for dengue diagnosis.

Kuno¹,

Cropp²,

Wong-Lee³

et al. 1998

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

show abstract

“…The samples were centrifuged and serum was separated and frozen at -20 o C until the serological analysis was performed. All serum samples were tested for the presence of anti-rubella IgM virus antibodies by using a commercial enzyme immunoassay (EIA) (Rubenostika IgM, Organon), for anti-measles virus IgM by using an antibody capture EIA developed at the Centers for Disease Control and Prevention (Atlanta, USA) (Hummel et al 1992), and for anti-dengue virus IgM by using an in-house EIA (Kuno et al 1987, Nogueira et al 1992. Those specimens, negative for rubella, measles and dengue virus antibodies, were also tested for anti-human parvovirus B19 IgM using an antibody capture EIA (MACEIA) (Cubel et al 1994, Nascimento et al 1998.…”

Section: Methodsmentioning

confidence: 99%

Clinical and epidemiological aspects of human parvovirus B19 infection in an urban area in Brazil (Niterói city area, State of Rio de Janeiro, Brazil)

Oliveira

Camacho

Pereira

et al. 2002

Mem. Inst. Oswaldo Cruz

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“…Serological results showed IgM seroconversion by IgM capture enzyme-linked immunosorbent assay (Kuno et al 1987) while G-ELISA (Miagostovich et al 1999) characterized dengue primary infection showing titers of <40 and 1/2560 on the acute and convalescent serum, respectively.…”

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confidence: 99%