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January 1, 2016Advances in Virology32 citationsOpen Access

Direct Detection and Identification of Enteroviruses from Faeces of Healthy Nigerian Children Using a Cell-Culture Independent RT-Seminested PCR Assay

TFTemitope O. C. FaleyeMAMoses Olubusuyi AdewumiBCBamidele Atinuke Coker

Key Result

A cell-culture independent RT-snPCR assay detected enteroviruses in 25% of faecal samples from healthy Nigerian children, revealing that species A enteroviruses are more commonly detected.

Study Design

Type

Cross-Sectional (n=60)

Structured PICO

P
Population
60 apparently healthy children in Ibadan, Nigeria
I
Intervention
Cell-culture independent RT-seminested PCR assay (WHO recommended protocol) for detection of enteroviruses from faecal samples
O
Outcome
Detection and identification of enterovirus diversity landscape

The cell-culture independent RT-snPCR protocol successfully detected a diverse range of enteroviruses, including species A enteroviruses, in healthy Nigerian children.

Abstract

Recently, a cell-culture independent protocol for detection of enteroviruses from clinical specimen was recommended by the WHO for surveillance alongside the previously established protocols. Here, we investigated whether this new protocol will show the same enterovirus diversity landscape as the established cell-culture dependent protocols. Faecal samples were collected from sixty apparently healthy children in Ibadan, Nigeria. Samples were resuspended in phosphate buffered saline, RNA was extracted, and the VP1 gene was amplified using WHO recommended RT-snPCR protocol. Amplicons were sequenced and sequences subjected to phylogenetic analysis. Fifteen (25%) of the 60 samples yielded the expected band size. Of the 15 amplicons sequenced, 12 were exploitable. The remaining 3 had electropherograms with multiple peaks and were unexploitable. Eleven of the 12 exploitable sequences were identified as Coxsackievirus A1 (CVA1), CVA3, CVA4, CVA8, CVA20, echovirus 32 (E32), enterovirus 71 (EV71), EVB80, and EVC99. Subsequently, the last exploitable sequence was identified as enterobacteriophage baseplate gene by nucleotide BLAST. The results of this study document the first description of molecular sequence data on CVA1, CVA8, and E32 strains present in Nigeria. The result further showed that species A enteroviruses were more commonly detected in the region when cell-culture bias is bypassed.

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Cite This Study

Faleye et al. (2016) conducted a cross-sectional in Healthy (n=60). Cell-culture independent RT-seminested PCR assay was evaluated on Detection of enteroviruses (expected band size). A cell-culture independent RT-snPCR assay detected enteroviruses in 25% of faecal samples from healthy Nigerian children, revealing that species A enteroviruses are more commonly detected.

synapsesocial.com/papers/6a16ef58c7240d1a707bcd02https://doi.org/10.1155/2016/1412838
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