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首页> 外文期刊>Indian journal of comparative microbiology, immunology and infectious dieases >MOLECULAR DETECTION AND QUANTIFICATION OF BLUETONGUE VIRUS IN SMALL RUMINANT AND CULICOIDES VECTORS IN GUJARAT, INDIA
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MOLECULAR DETECTION AND QUANTIFICATION OF BLUETONGUE VIRUS IN SMALL RUMINANT AND CULICOIDES VECTORS IN GUJARAT, INDIA

机译：印度古吉拉特邦小反刍动物和CULICOIDES载体中蓝舌病毒的分子检测和定量

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摘要

The present study was carried out to detect bluetongue virus (BTV) in Culicoides vectors and the biological samples of small ruminants with the use of PCR based molecular methods. The vectors were collected using the UV LED CDC traps, placed near animal sheds and then identified by morphological and PCR based methods. Primers specific to genus Culicoides and species Culicoides oxystoma were used in PCR reaction. The end point RT-PCR protocols were used for tentative and confirmatory diagnosis of BTV and its two serotypes viz. BTV-1 and BTV-16, whereas, the real time RT-PCR was used for confirmation of BTV and quantization of viral RNA load. Out of a total of 132 blood samples and 55 tissue samples viz., spleen, liver, kidney, thymus and heart blood of aborted fetuses and 09 pooled samples of Culicoides, total 21 (6.93%) samples i.e., 11 blood samples, 08 spleens and 02 pooled Culicoides samples were detected positive for BTV by NS1 gene specific RT-PCR. In nested PCR, 8 additional samples were found positive for BTV. Among these 29 positive samples, 6 were collected from spleens of aborted fetuses which was indicative of transplacental transmission of BTV among small ruminants. All the positive samples were of either of BTV-1 and BTV-16 serotypes as confirmed by serotype specific PCR. In real time PCR, plasmid vector based positive control was prepared, which confirmed the 21 samples to be BTV positive, whereas, quantization of viral RNA, showed that out of all clinical samples tested, highest and lowest RNA copy numbers i.e. 3566.41 and 81.79 were found in blood sample and spleen of sheep, respectively.
机译：本研究进行了检测在库蠓属向量和蓝舌病病毒(电视台)小反刍动物的生物样本基于PCR分子方法的使用。使用紫外线导致疾控中心向量收集陷阱,放在动物了被形态和PCR为基础方法。物种库蠓属oxystoma被用于PCR的反应。用于初步确认诊断北京电视台及其两种血清型即BTV-1 BTV-16,然而,用于实时rt - pcr确认北京电视台和病毒RNA的量子化负载。组织样本即、脾、肝、肾、胸腺和心脏的血流产胎儿和09年集中的样本库蠓属,共21例(6.93%)样品即11血液样本,08年脾脏和02集中库蠓属样本检测阳性北京电视台的NS1基因特定的rt - pcr。PCR 8额外样本发现阳性北京电视台。收集的流产胎儿脾脏表明经胎盘的传播吗北京电视台小反刍动物之一。样本的BTV-1 BTV-16血清型血清型特异性PCR证实了。实时PCR,质粒向量建立积极的控制了,这证实了21个样品是北京电视台积极,而量化病毒RNA,显示所有临床样品测试,最高及最低的RNA复制数字即3566.41和81.79中被发现血液样本和脾脏的绵羊。

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