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利用原位聚合酶链反应研究猪细小病毒2型基因组感染

Investigating porcine parvoviruses genogroup 2 infection using in situ polymerase chain reaction.

作者信息

Novosel Dinko, Cadar Daniel, Tuboly Tamás, Jungic Andreja, Stadejek Tomasz, Ait-Ali Tahar, Cságola Attila

机构信息

Department of Pathology, Croatian Veterinary Institute, Savska cesta 143, 10000, Zagreb, Croatia.

Department for Animal science, Faculty of Agriculture, University of Zagreb, Svetosimunska 25, 10000, Zagreb, Croatia.

出版信息

BMC Vet Res. 2018 May 21;14(1):163. doi: 10.1186/s12917-018-1487-z.

Abstract

BACKGROUND

Porcine parvovirus 2 (PPV2) was detected in swine serum without showing any relationship with disease. The emergence of the virus seemed to be a unique event until other genetically highly similar parvoviruses were identified in China and, later in 2012, the presence of the virus was also described in Europe. PPV2 is widely distributed in pig populations where it is suspected to be involved in respiratory conditions, based on its frequent detection in lung samples. In order to investigate the potential pathogenic involvement of PPV2, 60 dead pigs were examined from two farms. They were necropsied and tested for PPV2 and PCV2 (Porcine circovirus type 2) by PCR; by Brown and Brenn (B&B) staining for bacteria; by immunohistochemistry (IHC) to detect CD3, Swine leukocyte antigen class II DQ (SLAIIDQ), lysozyme, porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza (SIV), Mycoplasma hyopneumoniae (Mhyo); and by in situ hybridization (ISH) to detect ssDNA and dsDNA of PCV2. PPV2 positive samples were subjected to in situ polymerase chain reaction (IS-PCR) including double staining method to detect PPV2 and host cell markers. To calculate statistical difference we used GENMOD or LOGISTIC procedures in Statistical Analysis System (SAS®).

RESULTS

We found that the PPV2 was localized mostly in lymphocytes in lungs, lymph nodes and liver. Neither CD3 antigen nor lysozyme was expressed by these infected cells. In contrast, low levels of SLAIIDQ were expressed by infected cells, suggesting that PPV2 may have a specific tropism for immature B lymphocytes and/or NK lymphocytes though possibly not T lymphocytes.

CONCLUSION

The overall conclusion of this study indicates that PPV2 may contribute to the pathogenesis of pneumonia.

摘要

背景

在猪血清中检测到猪细小病毒2型(PPV2),且未发现其与疾病有任何关联。在其他基因高度相似的细小病毒在中国被鉴定出来之前,该病毒的出现似乎是一个独特事件;随后在2012年,欧洲也报道了该病毒的存在。PPV2在猪群中广泛分布,基于其在肺样本中的频繁检测,怀疑其与呼吸道疾病有关。为了研究PPV2的潜在致病作用,对来自两个猪场的60头死猪进行了检查。对它们进行了尸检,并通过聚合酶链反应(PCR)检测PPV2和猪圆环病毒2型(PCV2);通过布朗和布伦染色(B&B)检测细菌;通过免疫组织化学(IHC)检测CD3、猪白细胞抗原II类DQ(SLAIIDQ)、溶菌酶、猪繁殖与呼吸综合征病毒(PRRSV)、猪流感病毒(SIV)、猪肺炎支原体(Mhyo);通过原位杂交(ISH)检测PCV2的单链DNA(ssDNA)和双链DNA(dsDNA)。对PPV2阳性样本进行原位聚合酶链反应(IS-PCR),包括双重染色法以检测PPV2和宿主细胞标志物。为了计算统计学差异,我们在统计分析系统(SAS®)中使用了广义线性模型(GENMOD)或逻辑回归程序。

结果

我们发现PPV2主要定位于肺、淋巴结和肝脏的淋巴细胞中。这些被感染的细胞既不表达CD3抗原也不表达溶菌酶。相反,被感染的细胞表达低水平的SLAIIDQ,这表明PPV2可能对未成熟B淋巴细胞和/或自然杀伤细胞(NK细胞)具有特异性嗜性,尽管可能对T淋巴细胞没有嗜性。

结论

本研究的总体结论表明,PPV2可能参与肺炎的发病机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c84b/5963090/a104ee1618ef/12917_2018_1487_Fig1_HTML.jpg

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