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Isolation, Propagation and Genotyping of Human Rotaviruses Circulating among Children with Gastroenteritis in Two
Mona H El-Gayar1, Sarra E Saleh1, Aly F Mohamed2
1Department of Microbiology and Immunology, Faculty of Pharmacy, Ain Shams University, Organization of African Unity St., Cairo 11566, Egypt.
Insights
Group A rotavirus (RVA) causes infectious diarrhea in newborns. This study identified the G1P[8] genotype as predominant in Egyptian children, with Caco-2 cells proving effective for RVA propagation.
Area of Science:
- Virology
- Pediatrics
- Infectious Diseases
Background:
- Group A rotavirus (RVA) is a leading cause of infectious neonatal diarrhea globally.
- Limited data exists on RVA infection prevalence in Egyptian children.
Purpose of the Study:
- To isolate, propagate, and genotype human rotaviruses in Egyptian children with acute gastroenteritis.
- To assess the effectiveness of cell lines for RVA isolation and propagation.
Main Methods:
- Screening of 230 diarrheal samples using RT-PCR for RVA RNA.
- Genotyping of positive samples (n=34) using multiplex semi-nested PCR for G and P types.
- Propagation of RVA strains in Caco-2 and MA104 cell lines.
Main Results:
- RVA was detected in 14.8% (34/230) of samples.
- G1P[8] was the predominant genotype identified.
- Caco-2 cells demonstrated higher efficiency in RVA propagation compared to MA104 cells.
Conclusions:
- The G1P[8] rotavirus genotype circulates among Egyptian children with acute gastroenteritis.
- The Caco-2 cell line is a suitable and efficient model for propagating human rotaviruses from clinical samples.
Abstract:
The most prevalent cause of infectious neonatal diarrhea is Group A rotavirus (RVA). Unfortunately, there is a dearth of data on the incidence of rotavirus-associated infections among Egyptian children. The present study aimed to isolate, propagate, and genotype human rotaviruses circulating among Egyptian children with acute gastroenteritis admitted to two main university pediatric hospitals, Abo El-Reesh and El-Demerdash, over two consecutive winters, 2018-2020. Diarrheal samples (n = 230) were screened for Group A rotavirus RNA using RT-PCR assay. In positive samples (n = 34), multiplex semi-nested PCR was utilized to determine G and P genotypes. Thirty-four (14.8%) of the collected samples tested positive. The genotype distribution revealed that G1P[8] was the predominant rotavirus genotype throughout the current study. All rotavirus-positive fecal samples were passaged twice on human colorectal adenocarcinoma cell line (Caco-2) and rhesus monkey kidney epithelial cell line (MA104). Both cell lines could successfully isolate 14.7% (n = 5 out of 34) of the identified strains; however, Caco-2 cell line was shown to be more efficient than MA104 in promoting the propagation of human rotaviruses identified in Egyptian children's feces.
