COLLECTION OF THE STRAINS TESCHOVIRUS A, SAPELOVIRUS A, ENTEROVIRUS G OF THE INSTITUTE OF AGRICULTURAL MICROBIOLOGY AND AGROINDUSTRIAL MANUFACTURE OF THE NAAS

Derevianko S. V.
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Abstract

Objective. Arrangement of the collection of porcine enteroviruses (PEV) strains isolated on the territory of Ukraine in accordance with the requirements of the International Committee on Virus Taxonomy and supplementing it with new strains Teschovirus A (TV-A), Sapelovirus A (SV-A), Enterovirus G (EV-G). Methods. Virological, serological, molecular genetic, instrumental and statistical. Isolation, cultivation of viruses and determination of their biological activity were performed in passaged culture of porcine embryonic kidney cells (СНЕВ). The viral titre was calculated by the method of Reed and Muench. The typical affiliation of viruses was determined in the virus neutralization reaction in СНЕВ cell culture. Species affiliation was determined by reverse transcription polymerase chain reaction (RT PCR) using species-specific primers for TV-A, SV-A and EV-G, developed by us. Electron microscopy of viruses was performed on a transmission electron microscope by negative contrast enhancement method. Statistical processing was performed in Microsoft Office Excel and StatSoft STATISTICA 12. Results. As a result of the epizootic survey during 2002– 2019, 1,216 samples for virological testing were selected. Successive passages of СНЕВ cell culture resulted in obtaining 274 viral isolates. According to the results of studying physicochemical, morphological, biological properties of these isolated, they are classified as PEV. In connection with the change of taxonomy and nomenclature of PEV, serological and genetic reclassification of 30 strains of viruses isolated in Ukraine, including 14 reference strains according to the classification of V. P. Romanenko, 7 production strains, 9 strains with polyantigenic properties and 4 strains that did not have antigenic affinity with viruses of known PEV serotypes according to the classification of V. P. Romanenko was performed. It has been established that the reference strains of PEV according to the trivial classification of V. P. Romanenko belong to the species TV-A of the genus Teschovirus. As a result of conducted serological testing, PEV-10 M 2323, PEV-12 K 22, PEV-13 L 90, PEV-14 M 116, PEV-16 G 95, PEV-17 V 111, PEV-18 Ch 184, PEV-19 D 227, PEV-20 I 249, PEV-23 I 393 were classified as TV-A1; PEV-11 K 9, PEV-15 Ch 73 — as TV-A3, PEV-22 V 151 — as TV-A6. PEV-21 P 142 did not have antigen affinity with reference strains TV-A, SV-A and EV-G and belongs to a new serotype. Production strains of PEV-1 Perechynskyi 642, Bereznianskyi 652, Chernihivskyi 2372 were reclassified as TV-A1. PEV strains with polyantigenic properties such as G 31 and L 2661 have intertypic antigens with TV-A 1, 10, 11 and TV-A 3, 6, 10 serotypes, respectively. PEV strain of a new serotype Ch 881 was reclassified as SV-A. PEV strains Т 3, Ch 863, Ch 878 are the new serotypes of TV-A. Conclusion. As a result of studies, 274 viral isolates were isolated from 1,216 samples of material. The collection was supplemented with 20 reference strains of Teschovirus A, Sapelovirus A and Enterovirus G. Genetic and serological reclassification of 30 PEV strains isolated in Ukraine was performed. The collection of viral strains has been arranged in accordance with modern taxonomy and nomenclature. Seven viral strains were deposited. The collection of viruses has been supplemented with 4 strains of new serotypes of Teschovirus A.
收集了中国农业科学院农业微生物研究所和农业工业制造研究所的teschovirus a、sapelovirus a、肠病毒g株
目标。按照国际病毒分类学委员会的要求,安排在乌克兰境内分离的猪肠病毒(PEV)菌株的收集工作,并补充新菌株Teschovirus A (TV-A)、Sapelovirus A (SV-A)、Enterovirus G (EV-G)。方法。病毒学、血清学、分子遗传学、仪器学和统计学。在猪胚胎肾细胞传代培养中进行了病毒的分离、培养和生物活性测定(СНЕВ)。用Reed和Muench的方法计算病毒滴度。在СНЕВ细胞培养的病毒中和反应中确定了病毒的典型隶属关系。利用我们开发的TV-A、SV-A和EV-G的物种特异性引物,通过逆转录聚合酶链反应(RT - PCR)确定了物种归属。在透射电子显微镜上用负对比增强法对病毒进行电子显微镜观察。在Microsoft Office Excel和StatSoft STATISTICA 12中进行统计处理。结果。根据2002 - 2019年期间的动物流行病调查结果,选择了1216个样本进行病毒学检测。СНЕВ细胞培养连续传代,获得274株病毒分离株。根据对这些分离物的理化、形态学和生物学特性的研究结果,将其归类为PEV。结合PEV分类和命名法的变化,对乌克兰分离的30株病毒进行了血清学和遗传学的重新分类,其中14株为参考株,7株为生产株,9株具有多抗原特性,4株与已知PEV血清型病毒无抗原亲和力。根据罗曼年科病毒的琐碎分类,确定了PEV的参考菌株属于Teschovirus属的TV-A种。经血清学检测,将PEV-10 M 2323、PEV-12 K 22、PEV-13 L 90、PEV-14 M 116、PEV-16 G 95、PEV-17 V 111、PEV-18 Ch 184、PEV-19 D 227、PEV-20 I 249、PEV-23 I 393归为TV-A1;pev - 11k9, pev - 15ch73 -作为TV-A3, pev - 22v151 -作为TV-A6。pev - 21p142与参考菌株TV-A、SV-A和EV-G没有抗原亲和力,属于一种新的血清型。PEV-1生产菌株Perechynskyi 642、Bereznianskyi 652、Chernihivskyi 2372重新分类为TV-A1。具有多抗原特性的PEV菌株g31和l2661分别具有TV-A 1、10、11和TV-A 3、6、10血清型的型间抗原。将新血清型ch881的PEV毒株重新分类为SV-A型。PEV株Т 3、Ch 863、Ch 878是新的TV-A血清型。结论。研究结果表明,从1,216份材料样品中分离出274株病毒分离株。对乌克兰分离的30株PEV进行遗传和血清学重新分类。病毒株的收集已按照现代分类学和命名法进行了整理。共沉积了7株病毒株。收集的病毒中补充了4株新的甲型Teschovirus血清型。
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