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Molecular Detection of Human Enterovirus 71 Causing Hand, Foot and Mouth Disease in Klang Valley, Malaysia

Keywords: Cell culture , enterovirus 71 , hand foot and mouth disease , RT-PCR , sequencing

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Abstract:

Hand, Foot and Mouth Disease (HFMD) is a childhood infection caused by Enterovirus 71 (EV71) or Coxsackievirus A16 (CA16). Occasionally, Enterovirus 71 becomes fatal and causes encephalitis. The objective of the present study was to determine HFMD caused by Enterovirus 71 if prevalent in the Klang Valley, Malaysia. 184 specimens were collected from the patients reported to University Kebangsaan Malaysia Medical Centre (UKMMC). All the specimens were subjected to culture in rhabdomyosarcoma cell line and confirmed by Reverse transcription polymerase chain reaction (RT-PCR). The positive RT-PCR products were then sequenced to determine the viral genotype. Out of the 184 specimens, 89 showed cytopathic effects (CPE), indicating the presence of viral infections. Out of 89 positive CPE specimens 18 were positive with RT-PCR. Of the 18 positive specimens, 6 were Enterovirus 71, 3 Coxsackievirus A16, 8 Coxsackievirus A10, and 1 was vaccine-associated poliovirus 2. All the patients identified with strain Enterovirus 71 infection presented hand, foot and mouth disease and one of them had signs of paralysis as well. Collected Enterovirus 71 strains were classified under genotype C1 by phylogenetic analysis. This study proved that Enterovirus 71, genotype C1 prevalent in the study area and it did not cause serious outbreak in the Klang Valley, Malaysia. This prevalent strain could be used to choose for the development of a future vaccine candidate against HFMD.

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