Development and Validation of In-House Conventional and Multiplex PCR Methods for the Detection and Identification of Lophomonas spp.: An Innovative Approach

IF 2.9 4区医学 Q2 MEDICAL LABORATORY TECHNOLOGY

Journal of Clinical Laboratory Analysis Pub Date : 2025-05-08 DOI:10.1002/jcla.70049

Maryam Nakhaei, Mahdi Fakhar, Abouzar Bagheri, Hajar Ziaei Hezarjaribi, Saied Abediankenari, Ali Sharifpour, Maryam Ghasemi

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引用次数: 0

Abstract

Background

Pulmonary lophomoniasis is an emerging disease caused by the protozoan pathogen Lophomonas spp. Recently, a conventional polymerase chain reaction (PCR) method has been developed. However, its sensitivity and specificity remain to be fully established. Therefore, this study aimed to develop in-house conventional and multiplex PCR for the detection and identification of Lophomonas infections. Additionally, we attempted to compare the diagnostic performance of these novel PCR tests with the current microscopic examination method using BAL samples.

Methods

We studied 120 bronchoalveolar lavage (BAL) specimens of the patients clinically suspected of having lophomoniasis. The specimens were examined using three methods: microscopic examination (Giemsa staining), in-house conventional PCR, and multiplex-PCR. Moreover, multiplex-PCR was used for the simultaneous identification of two species of Lophomonas.

Results

Out of the 120 BAL specimens tested, 30 (25%) tested positive through microscopic wet mount examination. Among the three techniques, multiplex-PCR was the most sensitive (100%, 95% CI, 88.3–100), while Giemsa staining had the lowest sensitivity (86.2%, 95% CI, 69.4–94.5). The data reveal a strong agreement between multiplex-PCR and conventional PCR (κ = 0.96), while the lowest agreement was found between multiplex-PCR and microscopy methods (κ = 0.16). The study also confirmed the presence of L. blattarum species in all samples using multiplex-PCR.

Conclusions

This study demonstrates that the in-house multiplex-PCR is a robust and accurate diagnostic test for the detection and identification of Lophomonas species. Therefore, our findings suggest that this method may be a powerful tool to overcome some diagnostic pitfalls for lophomoniasis.

Abstract Image

查看原文本刊更多论文

开发和验证内部常规和多重PCR检测和鉴定Lophomonas spp.：一种创新的方法。

背景：肺吸虫病是由原生动物病原体吸虫单胞菌引起的一种新发疾病，近年来建立了传统的聚合酶链反应（PCR）方法。然而，其敏感性和特异性仍有待充分确定。因此，本研究旨在建立国内常规和多重PCR检测和鉴定Lophomonas感染。此外，我们试图将这些新型PCR检测的诊断性能与目前使用BAL样本的显微镜检查方法进行比较。方法：对120例临床疑似肺吸虫病患者的支气管肺泡灌洗（BAL）标本进行分析。标本检测采用三种方法：显微镜检查（吉姆萨染色），内部常规PCR和多重PCR。此外，采用多重pcr方法对两种Lophomonas进行了同时鉴定。结果：120例BAL标本中，30例（25%）湿片镜检阳性。三种技术中，多重pcr最敏感（100%,95% CI, 88.3-100），而吉姆萨染色最低（86.2%,95% CI, 69.4-94.5）。结果表明，多重聚合酶链式反应与常规PCR之间的一致性很强（κ = 0.96），而多重聚合酶链式反应与显微镜方法之间的一致性最低（κ = 0.16）。利用多重聚合酶链反应（multiple - pcr）证实了所有样品中均存在blattarum。结论：本研究表明，内部多重pcr是检测和鉴定Lophomonas的一种可靠、准确的诊断方法。因此，我们的研究结果表明，这种方法可能是一个强大的工具，以克服一些诊断陷阱的肺吸虫病。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Journal of Clinical Laboratory Analysis 医学-医学实验技术

CiteScore

5.60

自引率

7.40%

发文量

584

审稿时长

6-12 weeks

期刊介绍： Journal of Clinical Laboratory Analysis publishes original articles on newly developing modes of technology and laboratory assays, with emphasis on their application in current and future clinical laboratory testing. This includes reports from the following fields: immunochemistry and toxicology, hematology and hematopathology, immunopathology, molecular diagnostics, microbiology, genetic testing, immunohematology, and clinical chemistry.