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标准沙眼衣原体培养方法在进行一次额外的体外传代后敏感性得到提高。

Sensitivity of the Standard Chlamydia trachomatis Culture Method Is Improved After One Additional In Vitro Passage.

作者信息

Shao Lili, Guo Yuanli, Jiang Yong, Liu Yuanjun, Wang Mei, You Cong, Liu Quanzhong

机构信息

Department of Dermatovenereology, Tianjin Medical University General Hospital, Tianjin, China.

出版信息

J Clin Lab Anal. 2016 Sep;30(5):697-701. doi: 10.1002/jcla.21924. Epub 2016 Mar 14.

Abstract

BACKGROUND

Chlamydia trachomatis causes the most common bacterial sexually transmitted infection (STI) worldwide. Although highly sensitive nucleic acid amplification tests (NAATs) are used to routinely diagnose chlamydial infection, C. trachomatis isolation by cell culture is still preferred for legal cases and epidemiological studies because of its high specificity; however, the sensitivity of traditional two-passage diagnostic cultures is significantly lower than that of NAATs. Therefore, we sought to analyze if additional in vitro passaging of clinical samples would improve detection sensitivity of C. trachomatis.

METHODS

Clinical swabs (n = 428) were collected from Tianjin Medical University General Hospital, grown in McCoy cells for up to five passages, and analyzed for the presence of inclusions by iodine staining. Results were confirmed by routine PCR-based methods.

RESULTS

Viable C. trachomatis organisms were detected in 91 (21.26%) swabs with the traditional two-passage protocol, which increased to 145 (33.88%) and 149 (34.81%) following three and four passages, respectively. Thus, the standard protocol yielded a false-negative rate of nearly 39%. Subsequent PCR-based diagnostics revealed a concordance rate of 80.98% between these two methods without any false negatives.

CONCLUSION

The results of this study support the use of a three-passage Chlamydia culture procedure to increase the detection sensitivity of this method.

摘要

背景

沙眼衣原体导致全球最常见的细菌性性传播感染(STI)。尽管高灵敏度核酸扩增检测(NAATs)被用于常规诊断衣原体感染,但由于其高特异性,在法律案件和流行病学研究中,通过细胞培养分离沙眼衣原体仍更受青睐;然而,传统的两代诊断培养法的灵敏度显著低于NAATs。因此,我们试图分析临床样本额外的体外传代是否会提高沙眼衣原体的检测灵敏度。

方法

从天津医科大学总医院收集临床拭子(n = 428),在 McCoy 细胞中培养多达五代,并用碘染色分析包涵体的存在情况。结果通过基于常规 PCR 的方法进行确认。

结果

采用传统两代培养方案时,在 91 份(21.26%)拭子中检测到活的沙眼衣原体,在三代和四代传代后,分别增至 145 份(33.88%)和 149 份(34.81%)。因此,标准方案产生了近 39%的假阴性率。随后基于 PCR 的诊断显示这两种方法之间的一致率为 80.98%,且无假阴性。

结论

本研究结果支持采用三代衣原体培养程序来提高该方法的检测灵敏度。

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