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新鲜和冷冻牛粪中隐孢子虫卵囊的检测:三种方法的比较

Detection of Cryptosporidium oocysts in fresh and frozen cattle faeces: comparison of three methods.

作者信息

Brook E J, Christley R M, French N P, Hart C A

机构信息

Department of Veterinary Clinical Science, University of Liverpool, Leahurst, Neston, UK.

出版信息

Lett Appl Microbiol. 2008 Jan;46(1):26-31. doi: 10.1111/j.1472-765X.2007.02257.x. Epub 2007 Oct 15.

Abstract

AIMS

The aim of this study was to compare the performance of three commonly used screening tests for Cryptosporidium oocysts in fresh and frozen cattle faeces.

METHODS AND RESULTS

Twenty-nine freshly voided faecal samples were collected from calves from three farms in the northwest of England. Three diagnostic tests for Cryptosporidium were carried out on each sample both before and after freezing - the modified Ziehl-Neelsen (MZN) and auramine phenol (APh) stains and a commercial enzyme immunoassay (EIA) kit, the ProSpecT Cryptosporidium Microplate assay (Remel, Lenexa, KS). Twelve samples were deemed positive by the reference standard (polymerase chain reaction, PCR). There were some discrepancies between the results of the screening tests and the levels of agreement were quantified. The sensitivity and specificity of each method was determined, with PCR as the gold standard. Sensitivity and specificity of the MZN stain was optimized when samples with fewer than two oocyst-like bodies were classified as negative.

CONCLUSIONS

All three screening methods used were effective in detecting Cryptosporidium infection in both fresh and frozen calf faeces.

SIGNIFICANCE AND IMPACT OF THE STUDY

This study has highlighted the value of determining characteristics of tests used for diagnosis and epidemiological studies.

摘要

目的

本研究旨在比较三种常用于检测新鲜和冷冻牛粪便中隐孢子虫卵囊的筛查试验的性能。

方法与结果

从英格兰西北部三个农场的犊牛中收集了29份新鲜排出的粪便样本。对每个样本在冷冻前后都进行了三种隐孢子虫诊断试验——改良齐-尼氏(MZN)染色法、金胺酚(APh)染色法以及一种商业酶免疫分析(EIA)试剂盒,即ProSpecT隐孢子虫微孔板分析试剂盒(Remel公司,堪萨斯州莱尼克斯)。通过参考标准(聚合酶链反应,PCR)判定12个样本为阳性。筛查试验结果之间存在一些差异,并对一致性水平进行了量化。以PCR作为金标准,确定了每种方法的敏感性和特异性。当将少于两个卵囊样体的样本分类为阴性时,MZN染色法的敏感性和特异性得到了优化。

结论

所使用的三种筛查方法在检测新鲜和冷冻犊牛粪便中的隐孢子虫感染方面均有效。

研究的意义和影响

本研究突出了确定用于诊断和流行病学研究的检测方法特征的价值。

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