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通过聚合酶链反应检测到的柯萨奇病毒B3基因组:实验性小鼠心肌炎中心肌潜伏持续性的证据

Coxsackievirus B3 genomes detected by polymerase chain reaction: evidence of latent persistency in the myocardium in experimental murine myocarditis.

作者信息

Adachi K, Muraishi A, Seki Y, Yamaki K, Yoshizuka M

机构信息

Institute of Cardiovascular Diseases, Kurume University School of Medicine, Fukuoka, Japan.

出版信息

Histol Histopathol. 1996 Jul;11(3):587-96.

PMID:8839749
Abstract

We have investigated the time course after infection in coxsackievirus B3 murine myocarditis to determine the extent (if any) of persistent or latent infection that might be responsible for recurrence. We employed a polymerase chain reaction (PCR) method that can detect an extremely small amount of genome by amplification techniques in four-week-old BALD/c mice (n = 140) infected with coxsackievirus B3 by a single intraperitoneal injection of 1 x 10(4) plaque-forming units (PFU)/mouse (Group 1) and 1 x 10(2) PFU/mouse (Group 2). Mice were sacrificed on days 3, 5, 7, 10, 14, 21 and 28, and their hearts were resected for RNA extraction. Single chain DNA was synthesized from 1 microgram of RNA and the viral genome was amplified by PCR. The virus genome was strongly amplified in Group 1 from days 3 to 10, and in Group 2 from days 5 to 7, but afterwards both amplifications rapidly diminished. However, a positive signal, though very faint, persisted in both groups until day 28, by which time all histological evidence of myocarditis had disappeared in Group 2. Our results demonstrated that there was persistent or latent virus infection in the myocardium throughout the entire study period of 28 days. Such persistence might provide a pathomechanism for the exacerbation and recurrence of myocarditis.

摘要

我们研究了柯萨奇病毒B3型小鼠心肌炎感染后的时间进程,以确定可能导致复发的持续性或潜伏性感染的程度(如有)。我们采用了一种聚合酶链反应(PCR)方法,通过扩增技术在4周龄的BALB/c小鼠(n = 140)中检测极少量的基因组,这些小鼠通过腹腔注射1×10⁴空斑形成单位(PFU)/只(第1组)和1×10² PFU/只(第2组)感染柯萨奇病毒B3。在第3、5、7、10、14、21和28天处死小鼠,切除心脏用于RNA提取。从1微克RNA合成单链DNA,并通过PCR扩增病毒基因组。第1组在第3至10天病毒基因组强烈扩增,第2组在第5至7天强烈扩增,但之后两组的扩增均迅速减弱。然而,两组均有阳性信号持续存在,尽管非常微弱,直到第28天,此时第2组心肌炎的所有组织学证据均已消失。我们的结果表明,在整个28天的研究期间,心肌中存在持续性或潜伏性病毒感染。这种持续性可能为心肌炎的加重和复发提供一种发病机制。

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