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聚合酶链反应法早期检测实验感染小鼠血液中的迁移性旋毛虫幼虫。

Early detection by polymerase chain reaction of migratory Trichinella spiralis larvae in blood of experimentally infected mice.

机构信息

Department of Parasitology, Medical College, Zhengzhou University, Zhengzhou, People's Republic of China.

出版信息

Foodborne Pathog Dis. 2010 Aug;7(8):887-92. doi: 10.1089/fpd.2009.0472.

Abstract

We studied the sensitivity of polymerase chain reaction (PCR) for detecting DNA of migratory larvae of Trichinella spiralis at an early stage of infection with this parasite. We derived primers for PCR from a 1.6-kb repetitive sequence of the genome of T. spiralis and used PCR to detect Trichinella-specific DNA in blood of mice infected with 20, 100, or 300 muscle-derived larvae of T. spiralis at 3-21 days postinfection (dpi). We detected T. spiralis DNA in blood of mice infected with 20 larvae at 5 and 6 dpi, with a detection rate of 7.69% and in blood of mice infected with 100 larvae at 5-12 dpi, with a peak detection rate of 38.46% at 7 dpi. PCR detected T. spiralis larvae at 5-17 dpi in mice infected with 300 larvae, with detection rates exceeding 50% from 5 to 10 dpi and a peak rate of 61.54% at 7 dpi. The detection rates of T. spiralis larvae with PCR in the three groups of mice showed an increasing trend with an increase in the infecting dose of larval parasites (F = 17.811, p < 0.01). Our findings indicate that the sensitivity of PCR for detecting DNA migratory larvae of T. spiralis in blood of mice infected with this parasite depends on the severity of infection and the time elapsed after infection, and suggest that PCR may be useful for detecting Trichinella infection at an early stage in humans and food animals that test negatively for anti-Trichinella antibodies.

摘要

我们研究了聚合酶链反应(PCR)检测旋毛虫迁移幼虫 DNA 的敏感性,旋毛虫是一种寄生虫,处于感染的早期阶段。我们从旋毛虫基因组的 1.6kb 重复序列中推导了 PCR 引物,并使用 PCR 检测感染了 20、100 或 300 条肌肉来源的旋毛虫幼虫的小鼠血液中的旋毛虫特异性 DNA,感染后 3-21 天(dpi)。我们在感染后 5 和 6dpi 的感染 20 条幼虫的小鼠血液中检测到旋毛虫 DNA,检出率为 7.69%,在感染 100 条幼虫的小鼠血液中检测到 5-12dpi 的旋毛虫 DNA,检出率为 38.46%,峰值在 7dpi。在感染 300 条幼虫的小鼠中,PCR 在 5-17dpi 检测到旋毛虫幼虫,5-10dpi 的检出率超过 50%,7dpi 的峰值检出率为 61.54%。三组小鼠中旋毛虫幼虫 PCR 的检出率随着幼虫寄生虫感染剂量的增加而呈上升趋势(F=17.811,p<0.01)。我们的研究结果表明,PCR 检测感染旋毛虫的小鼠血液中 DNA 迁移幼虫的敏感性取决于感染的严重程度和感染后时间的长短,提示 PCR 可能有助于在人类和抗旋毛虫抗体检测阴性的食品动物中早期检测旋毛虫感染。

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