Isolation, propagation, and characterization of a G9P[4] human rotavirus strain 543765 in Iran

IF 2.4 3区医学 Q3 VIROLOGY

Virology Pub Date : 2025-09-17 DOI:10.1016/j.virol.2025.110695

Atefeh Kachooei , Reza Aramideh-Khouy , Mehrnaz Hosseini-Tehrani , Mahtab Mir-Hosseinian , Zahra Habib , Maryam Kazemi-Aghdam , Somayeh Jalivand , Tayebeh Latifi , Angila Ataei-Pirkooh , Zabihollah Shoja

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引用次数: 0

Abstract

Group A rotavirus (RVA) is a leading etiological agent of diarrheal diseases in children less than 5 years of age. While live attenuated RV vaccines have demonstrated high efficacy in high-income countries (HICs), their performance is substantially reduced in low- and middle-income countries (LMICs). Despite this disparity, the development and evaluation of live attenuated RVA strains remain a central objective in RV vaccine research. In this study, a human RVA strain, designated 543765, was successfully isolated from a stool sample using MA104 cell culture. The isolate was characterized through observation of cytopathic effects (CPE), polyacrylamide gel electrophoresis (PAGE), reverse transcription-polymerase chain reaction (RT-PCR), hemagglutination assay, transmission electron microscopy (TEM), and complete genome sequencing. Genotypic analysis revealed the following constellation: G9-P[4]-I1 (Lineage IV/II recombinant)-R1-C1-M1-A1-N1-T1-E1-H1. These findings suggest that strain 543765 exhibits stable structural and replication properties, achieving titers of up to 10⁸ TCID₅₀/mL in MA104 cells. Given its genetic profile and in vitro growth characteristics, strain 543765 holds promise as a candidate for development into a monovalent vaccine capable of inducing both homotypic and heterotypic immune protection against G9P[4] and other RVA genotypes. However, further investigation is warranted to evaluate whether serial passages in cell culture have resulted in attenuation, a determination that requires validation through clinical studies.

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伊朗g9p[4]人轮状病毒株543765的分离、繁殖和鉴定

A组轮状病毒（RVA）是5岁以下儿童腹泻病的主要病原。虽然RV减毒活疫苗在高收入国家（HICs）显示出很高的效力，但在低收入和中等收入国家（LMICs），其效力却大大降低。尽管存在这种差异，开发和评估RVA减毒活株仍然是RV疫苗研究的中心目标。在本研究中，使用MA104细胞培养成功地从粪便样本中分离出了一株命名为543765的人RVA菌株。通过观察细胞病变效应（CPE）、聚丙烯酰胺凝胶电泳（PAGE）、逆转录聚合酶链反应（RT-PCR）、血凝试验、透射电镜（TEM）和全基因组测序对分离物进行了鉴定。基因型分析显示：G9-P[4]-I1（谱系IV/II重组）-R1-C1-M1-A1-N1-T1-E1-H1。这些结果表明，菌株543765具有稳定的结构和复制特性，在MA104细胞中滴度高达108 TCID50/mL。鉴于其遗传特征和体外生长特性，菌株543765有望开发成一种单价疫苗，能够诱导针对G9P[4]和其他RVA基因型的同型和异型免疫保护。然而，需要进一步的研究来评估细胞培养中的连续传代是否会导致衰减，这需要通过临床研究来验证。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Virology 医学-病毒学

CiteScore

6.00

自引率

0.00%

发文量

157

审稿时长

50 days

期刊介绍： Launched in 1955, Virology is a broad and inclusive journal that welcomes submissions on all aspects of virology including plant, animal, microbial and human viruses. The journal publishes basic research as well as pre-clinical and clinical studies of vaccines, anti-viral drugs and their development, anti-viral therapies, and computational studies of virus infections. Any submission that is of broad interest to the community of virologists/vaccinologists and reporting scientifically accurate and valuable research will be considered for publication, including negative findings and multidisciplinary work.Virology is open to reviews, research manuscripts, short communication, registered reports as well as follow-up manuscripts.