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Prevalence and genetic heterogeneity of porcine group C rotaviruses in nursing and weaned piglets in Ohio USA and identification of a potential new VP4 genotype

机译:俄亥俄州俄亥俄州哺乳酸断奶仔猪豚鼠腐败的患病率和遗传异质性以及潜在的新VP4基因型的鉴定

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摘要

Swine fecal samples collected from seven farms were screened for group C rotaviruses (RVCs) using a reverse transcription-polymerase chain reaction assay. A total of 380 samples were tested and 19.5% were positive. Of the 128 samples collected in 2012, 23.5% from nursing piglets and 8.5% from weaned piglets were RVC positive, with a higher RVC frequency in diarrheic (28.4%) than in non-diarrheic (6.6%) piglets. Two strains (RVC/Pig-wt/USA/RV0104/2011/G3PX and RVC/Pig-wt/USA/RV0143/2012/G6Px) from two different farms were characterized genetically to gain information on virus diversity based on full length sequences of the inner capsid VP6, enterotoxin NSP4 and the outer capsid VP7 and VP4 (partial for RV0104) genes. The VP6 gene of the two strains showed high (99%) nucleotide identity to one another, 84–91% identity to other porcine RVCstrains and 81–82% identity to human and bovine RVC strains. The NSP4 gene analysis revealed that RVC/Pig-wt/USA/RV0104/2011/G3PX and RVC/Pig-wt/USA/RV0143/2012/G6Px strains were not closely related to each other (87% identity), but shared higher identity with prototype RVC/Pig-wt/USA/Cowden/1980/G1Px strain (93% and 89%, respectively) and were more distantly related to human strains (72–76% identity). The VP7 gene analysis indicated that the two strains were distantly related to one another (72% identity). RVC/Pig-wt/USA/RV0143/2012/G6Px was most closely related to porcine RVC G6 strains (82–86% identity), whereas RVC/Pig-wt/USA/RV0104/2011/G3PX was most closely related to porcine HF (G3) strain (94% identity). Analysis of the full length nucleotide sequence of the VP4 gene revealed that RVC/Pig-wt/USA/RV0143/2012/G6Px was distantly related to porcine (75%), bovine (74%) and human (70%) strains. The deduced amino acid identities (69.5–75.6%) of VP4 between RVC/Pig-wt/USA/RV0143/2012/G6Px and other RVCs were low; hence, we propose that this strain comprises a new VP4 genotype. Our results indicate high genetic heterogeneity in RVCs genes and the concurrent co-circulation of different genotypes at the same time. Our findings are useful for the development of more accurate diagnostic tools, for basic research to understand gene function and to provide information for RVC diversity germane to vaccine development.
机译:使用逆转录-聚合酶链反应试验,对从七个农场收集的猪粪样本进行C组轮状病毒(RVC)筛选。总共测试了380个样品,阳性率为19.5%。在2012年收集的128个样本中,护理仔猪的23.5%和断奶仔猪的8.5%为RVC阳性,腹泻(28.4%)的RVC频率高于非腹泻(6.6%)的猪。对来自两个不同农场的两种菌株(RVC / Pig-wt / USA / RV0104 / 2011 / G3PX和RVC / Pig-wt / USA / RV0143 / 2012 / G6Px)进行了遗传鉴定,以基于全长的序列获得病毒多样性信息。内衣壳VP6,肠毒素NSP4和外衣壳VP7和VP4(部分为RV0104)基因。两种菌株的VP6基因相互之间具有较高的核苷酸同一性(99%),与其他猪RVC菌株具有84-91%的同一性,与人和牛RVC菌株具有81-82%的同一性。 NSP4基因分析显示RVC / Pig-wt / USA / RV0104 / 2011 / G3PX和RVC / Pig-wt / USA / RV0143 / 2012 / G6Px菌株彼此之间没有密切关系(同一性为87%),但共享程度更高与原型RVC / Pig-wt / USA / Cowden / 1980 / G1Px菌株的同源性(分别为93%和89%),与人类菌株的亲缘关系更远(同一性为72-76%)。 VP7基因分析表明,这两个菌株彼此之间具有远距离关联(同一性为72%)。 RVC / Pig-wt / USA / RV0143 / 2012 / G6Px与猪RVC G6菌株最相关(同一性为82-86%),而RVC / Pig-wt / USA / RV0104 / 2011 / G3PX与猪最相关HF(G3)菌株(94%同一性)。 VP4基因的全长核苷酸序列分析表明,RVC / Pig-wt / USA / RV0143 / 2012 / G6Px与猪(75%),牛(74%)和人(70%)毒株关系密切。 RVC / Pig-wt / USA / RV0143 / 2012 / G6Px与其他RVC之间推定的VP4氨基酸同一性较低(69.5–75.6%);因此,我们建议该菌株包含新的VP4基因型。我们的结果表明RVCs基因具有高度的遗传异质性,并且同时存在着不同基因型的并发循环。我们的发现对开发更准确的诊断工具,基础研究以了解基因功能并为RVC多样性与疫苗开发的紧密联系提供信息很有用。

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