Xu Huirong, Xu Min, Chen Fang, Chen Hong, Du Wei, Yu Jing
Department of Ophthalmology, The Eighth Affiliated Hospital of Sun Yat-Sen University, Shenzhen, 518033, Guangdong Province, China.
Department of Ophthalmology, Subei People's Hospital affiliated to Yangzhou University, Yangzhou, 225001, Jiangsu Province, China.
BMC Ophthalmol. 2025 Jan 6;25(1):7. doi: 10.1186/s12886-025-03843-0.
This study aims to detect Mycobacterium tuberculosis complex (MTBC) DNA in intraocular fluid from clinically suspected tuberculous uveitis patients using multiplex polymerase chain reaction (PCR) and investigate the diagnostic utility of multiplex PCR for tuberculous uveitis.
Primers targeting three specific genes (MPB64, CYP141, and IS6110) within the MTBC genome were designed. Multiplex PCR was conducted using DNA from the H37Rv strain as well as DNA extracted from fluids of confirmed tuberculosis patients to assess primer specificity and method feasibility. Intraocular fluid samples were collected during the initial visit for multiplex PCR detection of MTBC DNA. The results of multiplex PCR tests were correlated with intraocular fluid findings and clinical profiles of patients clinically diagnosed with tuberculous uveitis who underwent standard antituberculosis therapy.
Multiplex PCR was employed to detect MTBC DNA in intraocular fluid samples from 15 patients clinically suspected of having tuberculous uveitis, with no amplification bands observed in the DNA lanes for the three target genes. T-cell spot test (T-SPOT) results were positive in 11 patients (100%), while purified protein derivative (PPD) tests were positive in 5 patients (45.5%). Abnormal chest CT findings were noted in 4 patients (36.4%), including one case of active pulmonary tuberculosis and three cases of inactive pulmonary tuberculosis. Retinal vasculitis was observed in 6 eyes (46.2%), panuveitis in 5 eyes (38.5%), and intermediate uveitis in 2 eyes (15.4%). The average duration of antituberculosis therapy administered to the 11 patients was 7.1 months (range: 6-10 months). The medium LogMAR Best Corrected Visual Acuity (BCVA) significantly improved at the last follow-up (Z=-2.371, P = 0.018).
Standard antituberculosis therapy demonstrated effectiveness in treating 11 patients clinically suspected of having tuberculous uveitis despite the absence of detectable MTBC DNA in intraocular fluid via multiplex PCR. Further investigation is warranted to elucidate the role of PCR in diagnosing ocular tuberculosis among Chinese individuals.
本研究旨在使用多重聚合酶链反应（PCR）检测临床疑似结核性葡萄膜炎患者眼内液中的结核分枝杆菌复合群（MTBC）DNA，并探讨多重PCR对结核性葡萄膜炎的诊断效用。
设计针对MTBC基因组中三个特定基因（MPB64、CYP141和IS6110）的引物。使用来自H37Rv菌株的DNA以及从确诊结核病患者的体液中提取的DNA进行多重PCR，以评估引物特异性和方法可行性。在初次就诊时收集眼内液样本，用于MTBC DNA的多重PCR检测。多重PCR检测结果与接受标准抗结核治疗的临床诊断为结核性葡萄膜炎患者的眼内液检查结果和临床资料相关。
采用多重PCR检测15例临床疑似结核性葡萄膜炎患者的眼内液样本中MTBC DNA，三个靶基因的DNA泳道均未观察到扩增条带。11例患者（100%）的T细胞斑点试验（T-SPOT）结果为阳性，5例患者（45.5%）的纯化蛋白衍生物（PPD）试验为阳性。4例患者（36.4%）胸部CT检查结果异常，包括1例活动性肺结核和3例非活动性肺结核。6只眼（46.2%）观察到视网膜血管炎，5只眼（38.5%）观察到全葡萄膜炎，2只眼（15.4%）观察到中间葡萄膜炎。11例患者接受抗结核治疗的平均疗程为7.1个月（范围：6-10个月）。末次随访时，平均最佳矫正视力（BCVA）的对数最小分辨角（LogMAR）显著改善（Z=-2.371，P = 0.018）。
尽管通过多重PCR在眼内液中未检测到可检测的MTBC DNA，但标准抗结核治疗对11例临床疑似结核性葡萄膜炎患者显示出有效性。有必要进一步研究以阐明PCR在中国人群眼结核诊断中的作用。
