Comparative detection efficacy of primers targeting SpeI-AvaI restriction fragment and small subunit ribosomal RNA gene of Babesia bigemina

IF 0.2 4区农林科学 Q4 AGRICULTURE, DAIRY & ANIMAL SCIENCE

Indian Journal of Animal Sciences Pub Date : 2023-08-02 DOI:10.56093/ijans.v92i7.109919

PARAMJIT KAUR, PRAYAG DUTT JUYAL, AMRITA SHARMA, LACHHMAN DAS SINGLA, CHANDRA SEKHAR MUKHOPADHYAY

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Abstract

The objective of the study is to evaluate the comparative detection efficacy of primers targeting SpeI-AvaI restriction fragment and small subunit ribosomal RNA (SSU rRNA) gene of Babesia bigemina by employing conventional polymerase chain reaction (PCR) on 783 animals (296 cattle and 487 buffaloes) of low lying (bet) area of Punjab. The detection rate of SpeI-AvaI and SSU rRNA PCR assays was 3.96% (31/783), and 6.64% (52/783), respectively. Among cattle and buffaloes, prevalence of B. bigemina was higher (P<0.01) in cattle by both the primers. The sensitivity and specificity of SSU rRNA PCR as compared to SpeI-AvaI restriction fragment PCR was 100% and 97.2%, respectively. The blast analysis of the nucleotides of the sequenced amplicons of Ludhiana isolates of SpeI-AvaI and SSU rRNA PCR assay of B. bigemina showed 83 and 100% similarity with available sequence in Genbank. The analysis of evolutionary divergence revealed that range of divergence was lying between 0.000 to 0.011 between SSU rRNA sequence with the other sequences of B. bigemina as well as Babesia species. To conclude, the primers targeting SSU rRNA gene are a better tool for amplification of the B. bigemina.

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双双巴贝斯虫SpeI-AvaI限制性内切片段与小亚基核糖体RNA基因引物检测效果比较

本研究采用常规聚合酶链式反应(PCR)方法，对印度旁遮普低洼地区783只动物(296头牛和487头水牛)进行双巴贝斯虫SpeI-AvaI限制性内切片段和SSU rRNA基因引物的检测效果进行比较。SpeI-AvaI和SSU rRNA PCR检测的检出率分别为3.96%(31/783)和6.64%(52/783)。在牛和水牛中，这两种引物对牛的感染率均较高(P<0.01)。与SpeI-AvaI限制性片段PCR相比，SSU rRNA PCR的灵敏度和特异性分别为100%和97.2%。对鲁地亚那分离株SpeI-AvaI序列扩增子核苷酸进行blast分析，对双芽孢杆菌进行SSU rRNA PCR检测，结果显示与Genbank中可用序列的相似性分别为83%和100%。进化差异分析表明，SSU rRNA序列与双芽贝虫和巴贝斯虫其他序列的差异范围在0.000 ~ 0.011之间。综上所述，以SSU rRNA基因为靶点的引物是一种较好的双叶双歧杆菌扩增工具。

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

Indian Journal of Animal Sciences 农林科学-奶制品与动物科学

CiteScore

0.60

自引率

25.00%

发文量

220

审稿时长

8 months

期刊介绍： Articles published in The Indian Journal of Animal Sciences encompass a broad range of research topics in animal health and production related to cattle, buffalo, sheep, goat, camel, equines, pig, rabbit, yak, mithun, poultry and fisheries. Studies involving wildlife species and laboratory animal species that address fundamental questions about their biology will also be considered for publication. All manuscripts must present some new development and must be original, timely, significant and scientifically excellent. Papers will be rejected if standards of care of, or procedures performed on animals are not up to those expected of humane veterinary scientists. At a minimum, standards must meet the International Guiding Principles for Biomedical Research involving Animals, as issued by the Council for International Organizations of Medical Sciences. (C.I.O.M.S., c/o WHO, CH 1211 Geneva 27, Switzerland). Articles reporting new animal disease must follow GOI directive as given in detail in Guidelines to Authors.