Detection of Norovirus by Detection of Norovirus by Enzyme Immunoassay (EIA) Enzyme Immunoassay (EIA)

in diarrhoeic stool of in diarrhoeic stool of hospitalized under five hospitalized under five

childrenchildren

Dr. Abul Kalam, Dr. S. K Paul, Dr. Abul Kalam, Dr. S. K Paul, Prof. Md. A. Hossain, M.U Ahmed, Prof. Md. A. Hossain, M.U Ahmed,

A.K.M Shamsuzzaman, C. A.K.M Shamsuzzaman, C. Mahmud, A.K.M. Musa, Mahmud, A.K.M. Musa,

WELCOMEWELCOME

TO THE PRESENTATION OFTO THE PRESENTATION OF Detection of Norovirus Detection of Norovirus

by Enzyme by Enzyme Immunoassay (EIA) in Immunoassay (EIA) in

diarrhoeic stool of diarrhoeic stool of hospitalized under five hospitalized under five

childrenchildren

Presented by Dr Abul KalamPresented by Dr Abul KalamM Phil (Thesis Part), Department of M Phil (Thesis Part), Department of Microbiology, MMCMicrobiology, MMC

Authors Authors Abul KalamAbul Kalam11, S. K Paul, S. K Paul11, Md. A. , Md. A.

HossainHossain11, M.U Ahmed, M.U Ahmed22, A.K.M , A.K.M ShamsuzzamanShamsuzzaman11, C. Mahmud, C. Mahmud11, , A.K.M. MusaA.K.M. Musa11, ,

• 1.Mymensingh medical college, 1.Mymensingh medical college,

• 2 Department of Medicine, faculty of 2 Department of Medicine, faculty of veterinary science , Bangladesh veterinary science , Bangladesh agricultural universityagricultural university

BackgroundBackground

• Gastroenteritis in the form of acute Gastroenteritis in the form of acute watery diarrhoea among under five watery diarrhoea among under five children remains a major public children remains a major public health problem throughout the world. health problem throughout the world.

• Gastroenteritis is caused by wide Gastroenteritis is caused by wide range of enteropathogens including range of enteropathogens including bacteria, viruses & protozoal agents.bacteria, viruses & protozoal agents.

• The viral causes of diarrhoea are The viral causes of diarrhoea are relatively less explored. relatively less explored.

Viral causes of Viral causes of gastroenteritisgastroenteritis

• Rotavirus, adenovirus, astrovirus and Rotavirus, adenovirus, astrovirus and Norovirus are well recognized causes Norovirus are well recognized causes of diarrhoea. of diarrhoea.

• Norovirus (NV) have appeared as one Norovirus (NV) have appeared as one of the most common causes of the most common causes epidemic gastroenteritis. epidemic gastroenteritis.

NorovirusNorovirus• Norovirus is heralded as the discovery Norovirus is heralded as the discovery

of the first virus specifically associated of the first virus specifically associated with gastroenteritis.with gastroenteritis.

• The The Norovirus Norovirus genus previously known genus previously known as Norwalk like virus (NLV), Small as Norwalk like virus (NLV), Small round structured virus (SRSV) is round structured virus (SRSV) is positive sense single stranded RNA positive sense single stranded RNA virus.virus.

• Norovirus is included in the family Norovirus is included in the family Caliciviridae. Caliciviridae.

Genogroups of NorovirusGenogroups of Norovirus

• Norovirus is Norovirus is divided into two divided into two distinct distinct genogroups, GGI genogroups, GGI and GGII, based on and GGII, based on genetic divergence genetic divergence in regions of the in regions of the RNA-dependent RNA-dependent RNA polymerase RNA polymerase and the major viral and the major viral capsid protein capsid protein (VP60).(VP60).

Genogroups Genogroups 11

GenogrouGenogroups2ps2

NorwalkNorwalk HawaiiHawaii

SouthampSouthamptonton

Snow Snow MountainMountain

ChibaChiba TorontoToronto

Desert Desert ShieldShield

Bristol Bristol virusesvirusesLordsdaleLordsdale

Research on NorovirusResearch on Norovirus• Research regarding Norovirus progressed Research regarding Norovirus progressed

slowly by being difficult to detect because slowly by being difficult to detect because of failure or difficulty in growing them in of failure or difficulty in growing them in cell culture, low levels of shedding in cell culture, low levels of shedding in feces, their small size, and the lack of feces, their small size, and the lack of suitable assays for their detection until suitable assays for their detection until 1992 when the development of molecular 1992 when the development of molecular biologic methods enabled rapid progress.biologic methods enabled rapid progress.

• Still vStill very little information is available ery little information is available regarding the impact of NV infection and regarding the impact of NV infection and prevalent genogroups in Bangladeshprevalent genogroups in Bangladesh

ObjectivesObjectives

• the present study was designed to the present study was designed to see the prevalence of Norovirus (NV) see the prevalence of Norovirus (NV) along with its genogroup in under along with its genogroup in under five children. five children.

Study place and periodStudy place and period

•This study was done in the This study was done in the department of Microbiology, department of Microbiology, Mymensingh Medical College Mymensingh Medical College

(in collaboration with the (in collaboration with the department of Medicine under department of Medicine under Faculty of veterinary science, Faculty of veterinary science, Bangladesh agricultural Bangladesh agricultural university)university)

•Study period was from March Study period was from March 2006 to February 20072006 to February 2007..

Subject selectionSubject selection

• A total of 92 stool samples obtained A total of 92 stool samples obtained from cases of acute watery diarrhoea from cases of acute watery diarrhoea admitted in paediatric ward of admitted in paediatric ward of Mymensingh Medical College Mymensingh Medical College Hospital.Hospital.

• Only under five children were Only under five children were selected for this study selected for this study

Methods and materialsMethods and materials

•The samples were tested by The samples were tested by Norovirus EIA (SRSV1 Norovirus EIA (SRSV1 TMTM, Senka , Senka Deiken Ltd ,Tokyo, Japan) Deiken Ltd ,Tokyo, Japan) according to kit manual.according to kit manual.

•The kit uses two wells for each The kit uses two wells for each specimen, one to detect specimen, one to detect Norovirus- genogroup (GI) and the Norovirus- genogroup (GI) and the other to detect Norovirus-other to detect Norovirus-genogroup (GII).genogroup (GII).

ResultResult

•Among the 92 stool Among the 92 stool samples, 4 (4.34%) were samples, 4 (4.34%) were positive for Norovirus. positive for Norovirus.

•All belong to Genogroup All belong to Genogroup 1. 1.

CommentComment

• This study showed that prevalence This study showed that prevalence of Norovirus is not remarkable in of Norovirus is not remarkable in Bangladeshi children Bangladeshi children

• This aspect needs further evaluation This aspect needs further evaluation including larger population using including larger population using molecular techniques like Reverse molecular techniques like Reverse transcriptase Polymerase chain transcriptase Polymerase chain reaction (RT- PCR). reaction (RT- PCR).