Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner-Schlossman syndrome in Chinese population
Digital droplet polymerase chain reaction analysis of common viruses in the aqueous humour of patients with Posner-Schlossman syndrome in Chinese population
复制标题
中国人群波斯纳-施洛斯曼综合征患者房水中常见病毒的数字液滴聚合酶链反应分析
DOI：
10.1111/ceo.13440
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发表时间：
2019-05-01
影响因子：
4
通讯作者：
Guan, Ming
中科院分区：
文献类型：
--
作者：
Cao, Guojun;Tan, Chen;Guan, Ming
目的比较定量聚合酶链反应（qPCR）和数字滴液聚合酶链反应（ddPCR）检测结果的一致性，确定ddPCR在检测房水中病毒的应用价值。方法收集60例Posner-Schlossman综合征（PSS）患者和70例老年性白内障患者的眼房水标本，并进行统计学分析。分别采用qPCR和ddPCR检测房水中人巨细胞病毒（HCMV）、单纯疱疹病毒（HSV）、EB病毒（EB病毒）和水痘带状疱疹病毒（水痘带状疱疹病毒）的靶核酸片段，用于PSS的诊断和疗效监测。结果不一致的样本通过下一代测序进行验证。结果ddPCR和qPCR分别检测出27例和20例病例组HCMV阳性。ddPCR将HCMV病毒检测的灵敏度从400拷贝/mL增加到100拷贝/mL。本研究未发现其他病原体。ddPCR结果与下一代测序结果一致。ddPCR法和qPCR法检测的Lg（HCMV拷贝数/mL）的平均值（SD）分别为1.66（1.92）和1.10（1.61）（P < 0.001）。与qPCR相比，ddPCR结果与临床疗效的一致性更好。所有ddPCR阳性的患者抗病毒治疗有效性良好，1个月内前房炎症缓解，角膜水肿消退，眼压控制良好。结论HCMV是中国人群中病原体引起的PSS的主要病因。ddPCR是一种有前途的工具，早期发现，准确诊断和治疗有效性监测病原体引起的PSS。ddPCR敏感性高，可避免重复前房穿刺。
Background To compare the detection results consistency of quantitative polymerase chain reaction (qPCR) and digital droplet polymerase chain reaction (ddPCR), and determine the value of ddPCR for viral detection in the aqueous humour. Methods A total of 130 aqueous humour samples were collected, including 60 patients with Posner-Schlossman syndrome (PSS) in case group and 70 elderly patients with senile cataract in control group. The target nucleic acid fragments of human cytomegalovirus (HCMV), herpes simplex virus, Epstein-Barr virus and varicella zoster virus in aqueous humour were analysed by qPCR and ddPCR, respectively, for the diagnosis and curative effect monitoring of pathogen-induced PSS. Samples with inconsistent results were verified by next-generation sequencing. Results There were 27 and 20 HCMV-positive cases detected in the case group by ddPCR and qPCR, respectively. ddPCR increased the sensitivity for the HCMV virus detection from 400 to 100 copies/mL. No other pathogens were found in this study. The results of ddPCR were consistent with that of next generation sequencing. The mean (SD) of Lg (HCMV copies/mL) detected by ddPCR and qPCR were 1.66 (1.92) and 1.10 (1.61), respectively (P < 0.001). Compared with qPCR, results of ddPCR showed better consistency with validity of clinical treatment. All patients with ddPCR-positive results had good validity on antiviral therapy, exhibiting anterior chamber inflammation remission, resolution of corneal oedema and good IOP control within 1 month. Conclusions HCMV was the leading cause of pathogen-induced PSS in the Chinese population. ddPCR was a promising tool for early detection, accurate diagnosis and therapeutic validity monitoring of pathogen-induced PSS. The high sensitivity of ddPCR could avoid repeated anterior chamber tap.