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Serotyping versus genotyping in infected sheep and goats with small ruminant lentiviruses
Veterinary Microbiology ( IF 3.3 ) Pub Date : 2020-11-20 , DOI: 10.1016/j.vetmic.2020.108931
Gabriel Eduardo Acevedo Jiménez 1 , Jorge Luis Tórtora Pérez 1 , Cecilia Rodríguez Murillo 1 , Beatriz Arellano Reynoso 2 , Hugo Ramírez Álvarez 1
Affiliation  

Despite SRLV infection being endemic in Mexico, there is little information regarding which genotypes are present. We compared serotyping and PCR-sequencing results from sheep and goats infected with SRLV. We separated plasma and peripheral blood leukocytes (PBL) from 1940 blood samples from sheep and goats from 12 states across Mexico. To detect SRLV infection, we tested plasma samples using two commercial ELISA kits (VMRD and Eradikit SRLV Screening). Then, we serotyped the infecting virus (A/ B) using Eradikit SRLV Genotyping. PBL DNA was used to detect the proviral genome via PCR. Positive amplicons were sequenced to identify viral genotypes using a phylogenetic analysis. Also, we analysed for residues differences in the sequences of a capsid epitope between genotypes. The serological results indicated a higher detection of seropositive animals using the VMRD ELISA compared to Eradikit, with 21 % and 15.3 % more in sheep and goats respectively. Only 25.7 % of the ELISA serotyping results matched those from PCR-sequencing. PCR-sequencing was able to identify genotype A, B and coinfections in animals classified as indeterminate by the ELISA test. This lack of sensitivity may be related to the lack of epitopes from the matrix and transmembrane peptides used by ELISA screening. Sequences analysis revealed that SRLVs found in sheep cluster with genetic subtypes A2 and B1, while those in goats cluster with subtypes A1 and B1. Serotyping did not prove to be an adequate method for predicting the viral genotype (A and / or B) in infections caused by SRLV.



中文翻译:

小反刍动物慢病毒感染绵羊和山羊的血清分型与基因分型

尽管在墨西哥是SRLV感染的地方病,但是关于存在哪些基因型的信息很少。我们比较了感染SRLV的绵羊和山羊的血清分型和PCR测序结果。我们从1940个来自墨西哥12个州的绵羊和山羊的血液样本中分离了血浆和外周血白细胞(PBL)。为了检测SRLV感染,我们使用两种商业ELISA试剂盒(VMRD和Eradikit SRLV筛选)测试了血浆样品。然后,我们使用Eradikit SRLV基因分型对感染病毒(A / B)进行血清分型。PBL DNA用于通过PCR检测原病毒基因组。使用系统发育分析对阳性扩增子进行测序,以鉴定病毒基因型。同样,我们分析了基因型之间衣壳表位序列中的残基差异。血清学结果表明,与Eradikit相比,使用VMRD ELISA对血清阳性动物的检出率更高,绵羊和山羊分别高21%和15.3%。ELISA血清分型结果中只有25.7%与PCR测序结果相符。PCR测序能够识别被ELISA测试归类为不确定的动物的基因型A,B和共感染。这种敏感性的缺乏可能与ELISA筛选所使用的基质和跨膜肽表位的缺乏有关。序列分析表明,在绵羊中发现具有遗传亚型A2和B1的SRLV,而在山羊中发现的SRLV则具有A1和B1亚型。血清分型并不能证明是预测由SRLV引起的感染中病毒基因型(A和/或B)的适当方法。绵羊和山羊分别增加3%。ELISA血清分型结果中只有25.7%与PCR测序结果相符。PCR测序能够识别被ELISA测试归类为不确定的动物的基因型A,B和共感染。这种敏感性的缺乏可能与ELISA筛选所使用的基质和跨膜肽表位的缺乏有关。序列分析表明,在绵羊中发现具有遗传亚型A2和B1的SRLV,而在山羊中发现的SRLV则具有A1和B1亚型。血清分型并不能证明是预测由SRLV引起的感染中病毒基因型(A和/或B)的适当方法。绵羊和山羊分别增加3%。ELISA血清分型结果中只有25.7%与PCR测序结果相符。PCR测序能够识别被ELISA测试归类为不确定的动物的基因型A,B和共感染。这种敏感性的缺乏可能与ELISA筛选所使用的基质和跨膜肽表位的缺乏有关。序列分析表明,在绵羊中发现具有遗传亚型A2和B1的SRLV,而在山羊中发现的SRLV则具有A1和B1亚型。血清分型并不能证明是预测由SRLV引起的感染中病毒基因型(A和/或B)的适当方法。ELISA测试将B和共感染归为不确定动物。这种敏感性的缺乏可能与ELISA筛选所使用的基质和跨膜肽表位的缺乏有关。序列分析显示,在绵羊中发现具有遗传亚型A2和B1的SRLV,而在山羊中发现具有个体A1和B1的SRLV。血清分型并未证明是预测由SRLV引起的感染中病毒基因型(A和/或B)的适当方法。ELISA测试将B和共感染归为不确定动物。这种敏感性的缺乏可能与ELISA筛选所使用的基质和跨膜肽表位的缺乏有关。序列分析显示,在绵羊中发现具有遗传亚型A2和B1的SRLV,而在山羊中发现具有个体A1和B1的SRLV。血清分型并不能证明是预测由SRLV引起的感染中病毒基因型(A和/或B)的适当方法。

更新日期:2020-12-01
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