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通过实时逆转录聚合酶链反应在患有猫传染性腹膜炎的猫中检测到的有和没有刺突基因突变的猫冠状病毒。

Feline coronavirus with and without spike gene mutations detected by real-time RT-PCRs in cats with feline infectious peritonitis.

作者信息

Emmler Laura, Felten Sandra, Matiasek Kaspar, Balzer Hans-Joerg, Pantchev Nikola, Leutenegger Christian, Hartmann Katrin

机构信息

Clinic of Small Animal Medicine, Centre for Clinical Veterinary Medicine, Ludwig Maximilian University of Munich, Munich, Germany.

Section of Clinical and Comparative Neuropathology, Institute of Veterinary Pathology, Centre for Clinical Veterinary Medicine, Ludwig Maximilian University of Munich, Munich, Germany.

出版信息

J Feline Med Surg. 2020 Aug;22(8):791-799. doi: 10.1177/1098612X19886671. Epub 2019 Nov 15.

Abstract

OBJECTIVES

Feline infectious peritonitis (FIP) emerges when feline coronaviruses (FCoVs) mutate within their host to a highly virulent biotype and the immune response is not able to control the infection. FCoV spike () gene mutations are considered to contribute to the change in virulence by enabling FCoV infection of and replication in macrophages. This study investigated the presence of FCoV with and without gene mutations in cats with FIP using two different real-time RT-PCRs on different samples obtained under clinical conditions.

METHODS

Fine-needle aspirates (FNAs) and incisional biopsies (IBs) of popliteal and mesenteric lymph nodes, liver, spleen, omentum and kidneys (each n = 20), EDTA blood (n = 13), buffy coat smears (n = 13), serum (n = 11), effusion (n = 14), cerebrospinal fluid (n = 16), aqueous humour (n = 20) and peritoneal lavage (n = 6) were obtained from 20 cats with FIP diagnosed by immunohistochemistry. Samples were examined by RT-PCR targeting the FCoV gene, detecting all FCoV, and gene mutation RT-PCR targeting mutations in nucleotides 23531 and 23537. The prevalence of FCoV detected in each sample type was calculated.

RESULTS

In 20/20 cats, FCoV with gene mutations was present in at least one sample, but there was variation in which sample was positive. FCoV with mutations in the gene was most frequently found in effusion (64%, 95% confidence interval [CI] 39-89), followed by spleen, omentum and kidney IBs (50%, 95% CI 28-72), mesenteric lymph node IBs and FNAs (45%, 95% CI 23-67), and FNAs of spleen and liver and liver IBs (40%, 95% CI 19-62).

CONCLUSIONS AND RELEVANCE

In these 20 cats with FIP, FCoVs with gene mutations were found in every cat in at least one tissue or fluid sample. This highlights the association between mutated gene and systemic FCoV spread. Examining a combination of different samples increased the probability of finding FCoV with the mutated gene.

摘要

目的

当猫冠状病毒(FCoV)在宿主体内突变为高毒力生物型且免疫反应无法控制感染时,猫传染性腹膜炎(FIP)就会出现。FCoV刺突()基因突变被认为通过使FCoV能够感染巨噬细胞并在其中复制,从而导致毒力变化。本研究使用两种不同的实时逆转录聚合酶链反应(RT-PCR),对临床条件下获取的不同样本进行检测,以调查患有FIP的猫中有无基因变异的FCoV的存在情况。

方法

从20只经免疫组化诊断为FIP的猫身上获取腘窝和肠系膜淋巴结、肝脏、脾脏、网膜和肾脏的细针穿刺抽吸物(FNA)和切开活检组织(IB)(各n = 20)、乙二胺四乙酸(EDTA)抗凝血(n = 13)、血沉棕黄层涂片(n = 13)、血清(n = 11)、积液(n = 14)、脑脊液(n = 16)、房水(n = 20)和腹腔灌洗物(n = 6)。通过针对FCoV基因的RT-PCR检测所有FCoV,并通过针对核苷酸23531和23537突变的基因突变RT-PCR对样本进行检测。计算每种样本类型中检测到的FCoV的患病率。

结果

在20只猫中,20只猫的至少一个样本中存在有基因突变的FCoV,但阳性样本存在差异。基因发生突变的FCoV最常见于积液(64%,95%置信区间[CI] 39 - 89),其次是脾脏、网膜和肾脏的IB(50%,95% CI 28 - 72)、肠系膜淋巴结的IB和FNA(45%,95% CI 23 - 67)以及脾脏和肝脏的FNA及肝脏的IB(40%,95% CI 19 - 62)。

结论及意义

在这20只患有FIP的猫中,每只猫的至少一个组织或液体样本中都发现了有基因突变的FCoV。这突出了基因变异与全身性FCoV传播之间的关联。检查不同样本的组合增加了发现有基因突变的FCoV的概率。

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本文引用的文献

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Transbound Emerg Dis. 2019 Mar;66(2):763-775. doi: 10.1111/tbed.13081. Epub 2018 Dec 5.
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Feline Coronavirus 3c Protein: A Candidate for a Virulence Marker?
Biomed Res Int. 2016;2016:8560691. doi: 10.1155/2016/8560691. Epub 2016 May 3.
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Investigation into the utility of an immunocytochemical assay in body cavity effusions for diagnosis of feline infectious peritonitis.
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Detection of feline coronavirus spike gene mutations as a tool to diagnose feline infectious peritonitis.
J Feline Med Surg. 2017 Apr;19(4):321-335. doi: 10.1177/1098612X15623824. Epub 2016 Jul 10.

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