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2.
J Infect Dis ; 214(12): 1897-1904, 2016 Dec 15.
Artigo em Inglês | MEDLINE | ID: mdl-27923950

RESUMO

BACKGROUND: Usually, immunoglobulin M (IgM) serologic analysis is not sufficiently specific to confirm Zika virus (ZIKV) infection. However, since IgM does not cross the placenta, it may be a good marker of infection in neonates. METHODS: We tested blood from 42 mothers and neonates with microcephaly and collected cerebrospinal fluid (CSF) specimens from 30 neonates. Molecular assays were performed for detection of ZIKV, dengue virus, and chikungunya virus; IgM enzyme-linked immunosorbent assays and plaque-reduction neutralization tests (PRNTs) were performed to detect ZIKV and dengue virus. No control neonates without microcephaly were evaluated. RESULTS: Among neonates, all 42 tested positive for ZIKV IgM: 38 of 42 serum specimens (90.5%) were positive, whereas 30 of 30 CSF specimens (100%) were positive. ZIKV IgM-specific ELISA ratios, calculated as the mean optical density (OD) of the test sample when reacted on viral antigen divided by the mean OD of the negative control when reacted with viral antigen, were higher in CSF specimens (median, 14.9 [range, 9.3-16.4]) than in serum (median, 8.9 [range, 2.1-20.6]; P = .0003). All ZIKV IgM-positive results among the neonates were confirmed by the detection of neutralizing antibodies. Mother/neonate pairs with primary ZIKV infection had neutralizing antibodies to ZIKV only, and mother/neonate pairs with ZIKV virus infection secondary to infection with another flavivirus had high titers of neutralizing antibodies to ZIKV. Among secondary infections, median titers in serum were 2072 (range, 232-12 980) for mothers and 2730 (range, 398-12 980) for neonates (P < .0001), and the median titer in CSF was 93 (range, 40-578) among neonates (P < .0001). CONCLUSIONS: Among neonates, detection of ZIKV IgM in serum is confirmatory of congenital ZIKV infection, and detection of ZIKV IgM in CSF is confirmatory of neurologic infection. Therefore, we recommend testing for ZIKV IgM in neonates suspected of having congenital ZIKV infection and performance of PRNTs in equivocal cases.


Assuntos
Anticorpos Neutralizantes/sangue , Anticorpos Neutralizantes/líquido cefalorraquidiano , Anticorpos Antivirais/sangue , Anticorpos Antivirais/líquido cefalorraquidiano , Imunoglobulina M/sangue , Infecção por Zika virus/diagnóstico , Zika virus/imunologia , Adolescente , Adulto , Sangue/imunologia , Líquido Cefalorraquidiano/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Recém-Nascido , Masculino , Testes de Neutralização , Sensibilidade e Especificidade , Ensaio de Placa Viral , Adulto Jovem , Infecção por Zika virus/congênito
3.
Sensors (Basel) ; 15(7): 15562-77, 2015 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-26140346

RESUMO

Dengue fever is the most prevalent vector-borne disease in the world, with nearly 100 million people infected every year. Early diagnosis and identification of the pathogen are crucial steps for the treatment and for prevention of the disease, mainly in areas where the co-circulation of different serotypes is common, increasing the outcome of dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS). Due to the lack of fast and inexpensive methods available for the identification of dengue serotypes, herein we report the development of an electrochemical DNA biosensor for the detection of sequences of dengue virus serotype 3 (DENV-3). DENV-3 probe was designed using bioinformatics software and differential pulse voltammetry (DPV) was used for electrochemical analysis. The results showed that a 22-m sequence was the best DNA probe for the identification of DENV-3. The optimum concentration of the DNA probe immobilized onto the electrode surface is 500 nM and a low detection limit of the system (3.09 nM). Moreover, this system allows selective detection of DENV-3 sequences in buffer and human serum solutions. Therefore, the application of DNA biosensors for diagnostics at the molecular level may contribute to future advances in the implementation of specific, effective and rapid detection methods for the diagnosis dengue viruses.


Assuntos
Técnicas Biossensoriais/métodos , DNA Viral/genética , Vírus da Dengue/genética , Sondas de DNA , DNA Viral/análise , DNA Viral/sangue , Dengue , Técnicas Eletroquímicas , Guanina , Humanos , Limite de Detecção , Oxirredução
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