Ananthakrishnan, P (2017) Prevalence and Molecular Characterisation of Circulating Serotypes of Rotavirus in Children less than 5 Years Old with Acute Watery Diarrhoea Attending Paediatric Department in a Tertiary Care Hospital. Masters thesis, Chengalpattu Medical College, Chengalpattu.
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Abstract
INTRODUCTION : Diarrheal diseases are one of the major reasons of illness and death in smaller children globally, particularly in less developed countries. Diarrhoea, especially acute watery diarrhoea, always a most important public health problem in all parts of the world. WHO estimated an attack rate of 3.2 episodes of diarrhoea per child every year globally. Diarrheal diseases result in about 2 million deaths every year, which makes it third among all causes of infectious disease deaths worldwide. Though global deaths from childhood diarrhoea decreased during the past 2 decades, there has been an increase in the number of hospitalizations due to diarrhoea. The decrease in diarrhoeal deaths may largely be attributed to the oral rehydration therapy, which has been reported to prevent 3 million dehydration deaths in a year globally. AIMS AND OBJECTIVES : 1. To estimate the prevalence of rota virus in smaller Children less than 5years of age attending paediatric department (Outpatient and In-patient) of Chengalpattu Medical College Hospital with acute watery diarrhoea 2. To find out the circulating serotypes of rotavirus in the confirmed cases 3. To do molecular characterisation of Rotavirus MATERIALS AND METHODS : Design of The Study: Cross sectional study Study Period: MAY 2015 TO APRIL 2016 Place of Study: Department of Microbiology, Chengalpattu Medical College, Chengalpattu Department of Paediatrics, Chengalpattu Medical College Hospital, Chengalpattu. King Institute of Preventive Medicine and Research Centre, Guindy, Chennai Selection Of Study Group: A total of 105 children, 0-5 years of age were included in the study. The clinical and demographic profiles obtained after obtaining proper consent from the parents / guardians. Inclusion Criteria : Children less than 5 years of age with complaints of more than three episodes of watery stools passed devoid of mucus and blood for a duration of less than 2 weeks and not on antibiotics or any laxatives were counted for this study. Exclusion Criteria : Children above 5 years of age Children with complaints of diarrhoea more than 2 weeks Children who had Rotavirus vaccine Children who acquired diarrhoeal illness after hospital admission were excluded from the study. Ethical Consideration : Ethical and research clearance was obtained from the Institutional Ethics Committee, Chengalpattu Medical College. Permission to conduct the study was sought from the Department of Paediatrics. Parents / guardians were enlightened about importance of the study and then consent was obtained from them before their child being incorporated into the study. Statistical Analysis : The statistical procedures were performed by the statistical package PASW (Predictive and Analysis Software) statistics -18 so called SPSS. The p values less than 0.05 were considered as significant. Collection and Transport of Stool Samples: The stool samples were collected in a clean wide mouthed sterile container, labelled properly, transported to the laboratory in ice pack for further processing. Samples were transported to The King Institute of Preventive Medicine, Guindy, Chennai, for PCR and further analysis. Throughout the transport and during preservation meticulous care was taken to ensure the cold chain of sample. Microbiological Analysis : A portion of each sample was stored in screw capped vials at -70ºc for analysis of Rota virus. Detection and Characterization of Rota Virus: Detection of Rota virus: All stool samples collected were subjected to Rotavirus identification tests with commercially available ELISA kit detecting group specific VP6 antigen for rota virus. 1. Antigen detection: was done by Enzyme Linked Immunosorbent assay method (ELISA) according to manufacturer’s instruction. [ROTACLONE] 2. Molecular characterisation and genotyping by conventional multiplex PCR. RESULTS : In this study 105 stool samples were collected from smaller children of less than 5 years of age, suffering from acute watery diarrhoea. And those are attending the paediatric Out-patient department or admitted for care as In-patients in Govt. Chengalpattu Medical College and Hospital. Rota virus antigen detection was done for all samples by ELISA. Out of 105 stool samples 24 showed positive results. Reverse Transcriptase Polymerase Chain Reaction was done for all the 24 ELISA positive samples for determination of G and P genotype of the rota virus stains. SUMMARY : This study was conducted at the Department of Microbiology; Chengalpattu Medical College and Hospital aimed at detection of Rotavirus in children with acute watery diarrhoea and Sero-tying. • Stools samples collected from 105 children with acute watery diarrhea were included in this study. • The samples were subjected to rotavirus antigen detection by VP6 specific monoclonal antibodies in Antigen Capture Enzyme Linked Immunosorbent Assay. • The samples which were positive for rotavirus were subjected to Reverse transcriptase polymerase chain reaction for characterisation of the rotavirus strains. • In the present study, most of the children with acute watery diarrhoea were in the age group 0-1 year, 55.23%, followed by 1-2 years, 30.47%. • Rotavirus prevalence was found to be 22.86% among children attending tertiary care hospital which was significantly associated with diarrhoea. • The main clinical presentation of rotavirus infected patients was acute watery diarrhoea associated with vomiting in 87.5% followed by fever in 79.17%. • Among ELISA positive cases 15 were found to be positive by RT-PCR. • 40% of the Rotavirus strains were fully typed for G and P Genotypes. 40% were partially typed and 20% were not typeable. • 80% of the G – Genotype and 40% of the P – Genotype could be determined. G9 Genotype was identified as predominant in G type, P[4] was identified as predominant P type. CONCLUSION : In the present study rotavirus was detected in 24 cases out of 105 children presented with acute watery diarrhoea. • Rotavirus prevalence was found to be 22.86% among children under 5 year of age with acute watery diarrhoea. • Majority of them were in the age group of 0-2 years. • The Genotype combinations of rotavirus identified is G9P[4], Partially typed – either G or P and GP both untypeable. • Thus the present study shows significant association of rotavirus in children under five years of age with acute watery diarrhoea and the prevalence of Serotype G9P4.The predominant rota virus strain in circulation appears to be G9P[4] in the study population, which is not a component of vaccines available at present. But the non typeable strains also pose an unknown antigenic challenge to the susceptible population. • Early diagnosis of Rotavirus in acute watery diarrhoea, continuous surveillance and prospective monitoring of its circulating strains and serotypes in any geographical area are important in public health perspective. It minimises the hospitalisation and mortality in children of under 5 age group. Any indiscriminate usage of antimicrobials in viral diarrhoea should be avoided and ORS should be encouraged. Effective vaccination prevents the disease burden and severity of Rotavirus in the community.
| Item Type: | Thesis (Masters) |
|---|---|
| Uncontrolled Keywords: | Prevalence ; Molecular Characterisation ; Circulating Serotypes ; Rotavirus ; Children less than 5 Years Old ; Acute Watery Diarrhoea ; Tertiary Care Hospital. |
| Subjects: | MEDICAL > Microbiology |
| Depositing User: | Subramani R |
| Date Deposited: | 21 Dec 2017 02:48 |
| Last Modified: | 21 Dec 2017 06:19 |
| URI: | http://repository-tnmgrmu.ac.in/id/eprint/4618 |
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