Nakhaei Maryam, Fakhar Mahdi, Bagheri Abouzar, Ziaei Hezarjaribi Hajar, Abediankenari Saied, Sharifpour Ali, Ghasemi Maryam
Toxoplasmosis Research Center, Communicable Diseases Institute, Mazandaran University of Medical Sciences, Sari, Iran.
Iranian National Registry Center for Lophomoniasis (INRCL), Imam Khomeini Hospital, Mazandaran University of Medical Sciences, Sari, Iran.
J Clin Lab Anal. 2025 Jun;39(12):e70049. doi: 10.1002/jcla.70049. Epub 2025 May 8.
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.
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.
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.
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.
肺嗜肺滴虫病是一种由原生动物病原体嗜肺滴虫属引起的新发疾病。最近，一种传统的聚合酶链反应（PCR）方法已经开发出来。然而，其敏感性和特异性仍有待充分确定。因此，本研究旨在开发用于检测和鉴定嗜肺滴虫感染的内部传统PCR和多重PCR。此外，我们试图将这些新型PCR检测方法的诊断性能与目前使用支气管肺泡灌洗（BAL）样本的显微镜检查方法进行比较。
我们研究了120例临床怀疑患有嗜肺滴虫病患者的支气管肺泡灌洗（BAL）标本。使用三种方法对标本进行检查：显微镜检查（吉姆萨染色）、内部传统PCR和多重PCR。此外，多重PCR用于同时鉴定两种嗜肺滴虫。
在检测的120份BAL标本中，30份（25%）通过显微镜湿片检查呈阳性。在这三种技术中，多重PCR最敏感（100%，95%CI，88.3 - 100），而吉姆萨染色的敏感性最低（86.2%，95%CI，69.4 - 94.5）。数据显示多重PCR与传统PCR之间有很强的一致性（κ = 0.96），而多重PCR与显微镜检查方法之间的一致性最低（κ = 0.16）。该研究还通过多重PCR证实了所有样本中均存在拟蠊嗜肺滴虫。
本研究表明，内部多重PCR是一种用于检测和鉴定嗜肺滴虫属的强大且准确的诊断测试。因此，我们的研究结果表明，该方法可能是克服嗜肺滴虫病某些诊断缺陷的有力工具。
