A comparison of virus isolation, immunohistochemistry, fetal serology, and reverse-transcription polymerise chain reaction assay for the identification of porcine reproductive and respiratory syndrome virus transplacental infection in the fetus
A comparison of virus isolation, immunohistochemistry, fetal serology, and reverse-transcription polymerise chain reaction assay for the identification of porcine reproductive and respiratory syndrome virus transplacental infection in the fetus
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DOI：
10.1177/104063870201400103
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发表时间：
2002-01-01
影响因子：
1.5
通讯作者：
Yoon, KJ
中科院分区：
文献类型：
--
作者：
Benson, JE;Yaeger, MJ;Yoon, KJ
对实验感染猪繁殖与呼吸综合征病毒（PRRSV）的母猪10窝共107胎进行病毒分离（VI）、免疫组化（IHC）、胎儿血清学和逆转录聚合酶链反应（RT-PCR）检测。除了比较每种检测的相对灵敏度和特异性外，还评价了RT-PCR作为样本的胸液和组织的相对适用性、自溶的影响以及合并胎仔样本的影响。免疫组化、免疫组化和胎儿血清学检测结果分别为48.6%、23.4%。和14.9%的107胎，并确定至少有1个感染的胎儿分别在10个，10个和5个10窝。子宫内死亡伴自溶降低了所有3种方法的检测有效性。胎仔胸腔液和组织同样适用于PRRSV的RT-PCR检测。当以1：1、1：2、1：4和1：8的稀释度进行评价时，来自VI阳性动物的胎仔组织或液体与来自阴性对照的相当材料合并对RT-PCR结果没有不利影响。在4、21或37 ℃下分别自溶24、48和96小时的VI阳性标本中，RT-PCR检测结果分别为100%、94.4%和83.3%。与其他检测方法相比，RT-PCR受自溶不良反应的影响最小。
Virus isolation (VI), immunohistochemistry (IHC), fetal serology, and reverse-transcription polymerise chain reaction assay (RT-PCR) were performed on samples from 107 fetuses comprising 10 litters taken from sows experimentally infected with porcine reproductive and respiratory syndrome virus (PRRSV). In addition to comparing the relative sensitivity and specificity of each test, RT-PCR was evaluated with respect to the relative suitability of thoracic fluids and tissues as samples, the effects of autolysis, and the effects of pooling of fetal specimens. VI, IHC, and fetal serology identified PRRSV infection in 48.6%, 23.4%. and 14.9% of 107 fetuses, respectively, and identified at least 1 infected fetus in 10, 10, and 5 of 10 litters, respectively. In utero death with autolysis reduced the test efficacy of all 3 methods. Fetal thoracic fluid and tissues were equally suitable for RT-PCR detection of PRRSV Pooling fetal tissues or fluids from VI-positive animals with comparable material from negative controls had no detrimental effect on RT-PCR results when evaluated at dilutions of 1:1, 1:2, 1:4, and 1:8. The results of RT-PCR testing were positive in 100%, 94.4%, and 83.3% of VI-positive specimens allowed to autolyze at 4, 21, or 37 C, respectively, for 24, 48, and 96 hours. Compared with the other testing modalities, RT-PCR appeared to be impacted the least by the adverse effects of autolysis.