Immunological and molecular detection of rotavirus genotype in calves with gastroenteritis in Diyala-Iraq

Ammar Talib Nasser, A. Hasan, Amer Khazaal Saleh, M. Saleh
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Abstract

Aim: To explore the prevalence of rotavirus infection along with the molecular detection and genotyping of group A rotavirus (RVA) among bovine calves up to 5 months old in Diyala province-Iraq. Methods: This is a cross sectional study conducted in Diyala province-Iraq during the period of 2019-2020. One hundred bovine calves with age range of 1-5 months were included in the study. All were suffering acute gastroenteritis. Serum anti-rotavirus IgM and IgG plus fecal rotavirus Ag were tested for using ELISA techniques. Stool samples positive for rotavirus Ag were submitted for reverse transcription PCR (RT-PCR) for G and P genes, followed by sequencing and genotyping thereafter. Statistical analysis was done using SPSS version 25 and P values ≤ 0.05 were considered significant. Results: The positivity rate of anti-rotavirus IgM was 80% (P = 0.0001), and that of anti-rotavirus IgG was 79% (P = 0.0001). The rotavirus stool antigen was detected in 68% of calves (P = 0.01). A total of 45 stool samples which were positive for rotavirus Ag were submitted for RT-PCR; 13 (28.9%) were positive and 32 (71.1%) were negative (P = 0.084). 10 PCR positive samples were used for sequencing and genotyping and indicated that all investigated strains belonged to G1P[8] genotype. Conclusion: The current strains analyzed belonged to the G1P[8] RVA genotypes, affirming that employment of VP7 gene polymorphism accurately yielded uniform phylogenetic distances amongst investigated rotavirus strains and that there were no noticeable assortment events between human and animal rotavirus strains in Diyala province.
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伊拉克迪亚拉地区犊牛胃肠炎轮状病毒基因型的免疫学和分子检测
目的:了解伊拉克迪亚拉省5月龄以下犊牛中轮状病毒感染的流行情况、A组轮状病毒(RVA)的分子检测和基因分型。方法:采用横断面研究方法,于2019-2020年在伊拉克迪亚拉省开展。研究对象为100头年龄在1 ~ 5月龄的小牛。所有人都患有急性肠胃炎。采用ELISA技术检测血清抗轮状病毒IgM和IgG +粪便轮状病毒Ag。将轮状病毒Ag阳性的粪便样本进行G和P基因的反转录PCR (RT-PCR),然后进行测序和基因分型。采用SPSS 25进行统计学分析,P值≤0.05为差异有统计学意义。结果:抗轮状病毒IgM阳性率为80% (P = 0.0001),抗轮状病毒IgG阳性率为79% (P = 0.0001)。轮状病毒粪便抗原检出率为68% (P = 0.01)。对45份轮状病毒抗原阳性的粪便标本进行RT-PCR检测;阳性13例(28.9%),阴性32例(71.1%)(P = 0.084)。10份PCR阳性样本进行测序和基因分型,结果表明所调查的菌株均为G1P[8]基因型。结论:目前分析的毒株属于G1P[8] RVA基因型,证实利用VP7基因多态性准确地获得了在所调查的轮状病毒毒株之间统一的系统发育距离,并且在迪亚拉省没有明显的人与动物轮状病毒毒株之间的配种事件。
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