Sensitivity of RT-PCR method in samples shown to be positive for Zika virus by RT-qPCR in vector competence studies

Paiva, Marcelo Henrique Santos; Guedes, Duschinka Ribeiro Duarte; Leal, Walter Soares; Ayres, Constância Flávia Junqueira

doi:10.1590/1678-4685-gmb-2016-0312

Cited by 19 publications

(18 citation statements)

References 14 publications

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“…To perform the real-time PCR we used the QuantStudio 3 Real-Time PCR System (Thermo Fisher Scientific ®) and the cut-off Ct value was ≤39 based on different criteria, among which are CDC interpretation criteria, considering a specimen positive if primer sets showed amplification with cycle threshold (CT) values ≤38.5 [ 31 ]. Different papers have showed that Zika virus could be tested positive in human and mosquito samples with CT values of 38–39 [ 32 – 34 ]; and a recent paper from Salvador, Bahia, Brazil, showed that serum samples with CTs > 34 could be confirmed positive for sequencing confirming that low viral copies are common in Zika virus positive patients [ 35 , 36 ]. As positive control of ZIKV we used an isolated virus from Aedes aegypti, which was donated by the Programa de Estudio y Control de Enfermedades Tropicales (PECET) de la Universidad de Antioquia and as negative control we used RNAse free water.…”

Section: Methodsmentioning

confidence: 99%

Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border

Carrillo-Hernández

Ruíz-Saenz

Villamizar³

et al. 2018

BMC Infect Dis

140

124

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BackgroundIn Colombia, the dengue virus (DENV) has been endemic for decades, and with the recent entry of the chikungunya virus (CHIKV) (2014) and the Zika virus (ZIKV) (2015), health systems are overloaded because the diagnosis of these three diseases is based on clinical symptoms, and the three diseases share a symptomatology of febrile syndrome. Thus, the objective of this study was to use molecular methods to identify their co-circulation as well as the prevalence of co-infections, in a cohort of patients at the Colombian-Venezuelan border.MethodsA total of 157 serum samples from patients with febrile syndrome consistent with DENV were collected after informed consent and processed for the identification of DENV (conventional PCR and real-time PCR), CHIKV (conventional PCR), and ZIKV (real-time PCR). DENV-positive samples were serotyped, and some of those positive for DENV and CHIKV were sequenced.ResultsEighty-two patients were positive for one or more viruses: 33 (21.02%) for DENV, 47 (29.94%) for CHIKV, and 29 (18.47%) for ZIKV. The mean age range of the infected population was statistically higher in the patients infected with ZIKV (29.72 years) than in those infected with DENV or CHIKV (21.09 years). Both co-circulation and co-infection of these three viruses was found. The prevalence of DENV/CHIKV, DENV/ZIKV, and CHIKV/ZIKV co-infection was 7.64%, 6.37%, and 5.10%, with attack rates of 14.90, 12.42, and 9.93 cases per 100,000 inhabitants, respectively. Furthermore, three patients were found to be co-infected with all three viruses (prevalence of 1.91%), with an attack rate of 4.96 cases per 100,000 inhabitants.ConclusionOur results demonstrate the simultaneous co-circulation of DENV, CHIKV, ZIKV and their co-infections at the Colombian-Venezuelan border. Moreover, it is necessary to improve the differential diagnosis in patients with acute febrile syndrome and to study the possible consequences of this epidemiological overview of the clinical outcomes of these diseases in endemic regions.Electronic supplementary materialThe online version of this article (10.1186/s12879-018-2976-1) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border

Carrillo-Hernández

Ruíz-Saenz

Villamizar³

et al. 2018

BMC Infect Dis

140

124

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“…However, the diagnosis of this infection poses some difficulties due to the possible cross-reaction with other flaviviruses. 5 In terms of the neuroimaging findings, a variety of cortical malformations can be identified, including parenchymal volume reduction, alterations in the gyral pattern as well in white and gray matter, ventriculomegaly, abnormalities of the corpus callosum and cerebellum, hypoplasia and/ or atrophy of the brain stem, and diffuse calcifications. 6 In 2016, the World Health Organization reported cases of microcephaly and neurological disorders associated with ZIKV as an international public-health emergency, thereby intensifying the need for further research in this area.…”

Section: Introductionmentioning

confidence: 99%

Review of neuroimaging findings in congenital Zika virus syndrome and its relation to the time of infection

et al. 2020

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Background Many original articles and case series have been published emphasizing the neuroimaging findings of congenital Zika virus (ZIKV) infection. The majority of these studies do not follow a neuroradiological methodology to describe malformations and brain abnormalities resulting from ZIKV infection. The cause-and-effect correlation between the gestational period of maternal infection and the severity of encephalic changes at birth has rarely been reported. A systematic literature review was conducted on the neuroimaging findings in children affected with microcephaly due to ZIKV. Methods PubMed, Cochrane Library and Web of Science were searched for full-text articles published up to July 2019. Duplicate entries were removed. Two independent reviewers performed a quality assessment of all the studies included. Results A total of 2214 publications were identified. Of these 2170 were excluded by analysis of titles and abstracts, resulting in the inclusion of only eight articles. Chi-square and Fisher’s exact tests were performed with a 95% confidence interval to verify the statistically significant differences in the neuroradiological findings between the cases of ZIKV infection in the first or second trimester of gestation. The studies published so far have described image abnormalities at random, without utilizing any pre-established neuroradiological criteria, and imaging modalities with different sensitivity and accuracy have been used, which jeopardizes a reliable and adequate statistical analysis. Conclusions Neuroimaging abnormalities are much more prevalent and severe when the infection by ZIKV is contracted in the first or second trimester of pregnancy.

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“…Previous studies reported that conventional RT-PCR could detect ZIKV viral load in most of the samples, with an estimated 10 6 -10 10 RNA copies/mL. 23,30 Published studies showed strong association between microcephaly and ZIKV infection confirmed by qRT-PCR, capture IgM ELISA, or both. 21,31,32 In Brazil, 32 of 91 neonates born with microcephaly were confirmed positive for ZIKV infection by qRT-PCR or anti-ZIKV IgM ELISA with confirmation more frequent in CSF than in serum.…”

Section: Discussionmentioning

confidence: 99%

“…Previous studies reported that conventional RT-PCR could detect ZIKV viral load in most of the samples, with an estimated 10 6 –10 10 RNA copies/mL. 23 , 30 …”

Section: Discussionmentioning

confidence: 99%

Congenital Zika Virus Infection in a Birth Cohort in Vietnam, 2017–2018

Tun

Moriuchi

Toizumi

et al. 2020

The American Journal of Tropical Medicine and Hygiene

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To detect congenital ZIKV infection (CZI) in a birth cohort and among high-risk neonates in Vietnam, we collected umbilical cord blood plasma samples of newly delivered babies and peripheral plasma samples of high-risk neonates in Nha Trang, central Vietnam, between July 2017 and September 2018. Samples were subjected to serological and molecular tests. Of the 2013 newly delivered babies, 21 (1%) were positive for Zika virus (ZIKV) IgM and 1,599 (79%) for Flavivirus IgG. Among the 21 ZIKV IgM-positives, 11 were confirmed to have CZI because their plasma samples had anti-ZIKV neutralization titers ³ 4 times higher than those against dengue virus (DENV)-1 to 4 and Japanese encephalitis virus (JEV) and were tested for the ZIKV RNA positive by real-time reverse transcription-PCR. Therefore, the incidence of CZI in our birth cohort was approximately 0.5%. Of the 150 high-risk neonates, three (2%) and 95 (63%) were positive for ZIKV IgM and Flavivirus IgG antibodies, respectively. None of the three ZIKV IgM-positives had ³ 4 times higher anti-ZIKV neutralization titers than those against DENV-1 to 4 and JEV, and were therefore considered as probable CZI. Our results indicate that CZI is not rare in Vietnam. Although those with confirmed CZI did not show apparent symptoms suspected of congenital Zika syndrome at birth, detailed examinations and follow-up studies are needed to clarify the CZI impact in Vietnam. This is the first report of CZI cases in a birth cohort in Asia.

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Sensitivity of RT-PCR method in samples shown to be positive for Zika virus by RT-qPCR in vector competence studies

Cited by 19 publications

References 14 publications

Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border

Co-circulation and simultaneous co-infection of dengue, chikungunya, and zika viruses in patients with febrile syndrome at the Colombian-Venezuelan border

Review of neuroimaging findings in congenital Zika virus syndrome and its relation to the time of infection

Congenital Zika Virus Infection in a Birth Cohort in Vietnam, 2017–2018

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