Detection of Leptospira spp. using polymerase chain reaction technique from kidney of Rattus norvegicus from Grenada, West Indies

Q2 Veterinary
Bhumika K. Sharma, Katelyn Thille, Nia Rametta, Ravindra Sharma
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Abstract

Aim: This study aimed to find out the prevalence of active infection of Leptospira spp. in Rattus norvegicus from Grenada, West Indies, through polymerase chain reaction (PCR). Materials and Methods: One hundred and forty-nine rats were trapped, anesthetized and their kidneys collected aseptically. DNA was extracted from the kidney tissue of each rat. PCR was performed targeting LipL32 gene. Eighteen PCR-positive amplicons for LipL32 gene segment were purified and sent for direct sequencing to the sequencing facility of MCLAB (South San Francisco, USA). Results of sequencing were read and interpreted. The prevalence of Leptospira spp. in relation to sex and age was also recorded. Results: All amplified sequences were compared to the sequences present in GenBank using basic local alignment search tool (BLAST) from the online website National Center for Biotechnology Information, the results revealed that six samples had similarity to Leptospira interrogans strain 1399/2016 and eight samples had similarity with Leptospira borgpetersenii serovar Hardjo-bovis strain L49. Of 149 kidney samples, only 14 were positive for Leptospira spp. by PCR giving an incidence of 9.3%. There was no significant difference found in relation to sex and age. Conclusion: This is the first report confirming active infection of Leptospira spp. in Rattus norvegicus in Grenada using PCR. The presence of active infection in rats can be considered as high risk for humans. Further research to understand the epidemiology of leptospirosis in Grenada is suggested.
应用聚合酶链反应技术检测西印度群岛格林纳达褐家鼠肾脏钩端螺旋体
目的:本研究旨在通过聚合酶链式反应(PCR)了解西印度群岛格林纳达褐家鼠钩端螺旋体的活动感染率。材料和方法:将一百四十九只大鼠捕获、麻醉并无菌收集肾脏。从每只大鼠的肾组织中提取DNA。针对LipL32基因进行PCR。纯化LipL32基因片段的18个PCR阳性扩增子,并将其送至MCLAB(美国南旧金山)的测序设施进行直接测序。读取并解释测序结果。还记录了钩端螺旋体的流行率与性别和年龄的关系。结果:使用国家生物技术信息中心的基本局部比对搜索工具(BLAST)将所有扩增的序列与GenBank中的序列进行比较,结果显示,6个样本与询问型钩端螺旋体1399/2016菌株相似,8个样本与博格彼得森细端螺旋体血清型Hardjo bovis菌株L49相似。在149份肾脏样本中,只有14份钩端螺旋体阳性。通过PCR检测,发病率为9.3%。性别和年龄之间没有显著差异。结论:这是首次用聚合酶链式反应证实格林纳达褐家鼠钩端螺旋体的活动性感染。大鼠存在活动性感染可被认为是人类的高风险。建议进一步研究以了解格林纳达钩端螺旋体病的流行病学。
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来源期刊
International Journal of One Health
International Journal of One Health Veterinary-Veterinary (all)
CiteScore
2.10
自引率
0.00%
发文量
13
审稿时长
15 weeks
期刊介绍: International Journal of One Health publishes high quality and novelty papers focusing on One Health. Review articles are highly appreciated. All articles published by International Journal of One Health are made freely and permanently accessible online. All articles to International Journal of One Health are posted online immediately as they are ready for publication.
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