评估一种多重聚合酶链反应检测法用于检测溃疡性结肠炎患者粪便样本中的巨细胞病毒。
Evaluation of a multiplex PCR assay for detection of cytomegalovirus in stool samples from patients with ulcerative colitis.
作者信息
Nahar Saifun, Iraha Atsushi, Hokama Akira, Uehara Ayako, Parrott Gretchen, Ohira Tetsuya, Kaida Masatoshi, Kinjo Tetsu, Kinjo Takeshi, Hirata Tetsuo, Kinjo Nagisa, Fujita Jiro
机构信息
Saifun Nahar, Ayako Uehara, Gretchen Parrott, Tetsu Kinjo, Takeshi Kinjo, Tetsuo Hirata, Jiro Fujita, Department of Infectious, Respiratory, and Digestive Medicine, University of the Ryukyus, Okinawa 903-0215, Japan.
出版信息
World J Gastroenterol. 2015 Nov 28;21(44):12667-75. doi: 10.3748/wjg.v21.i44.12667.
AIM
To evaluate a multiplex PCR assay for the detection of bacterial and viral enteropathogens in stool samples from patients with ulcerative colitis (UC).
METHODS
We prospectively analyzed 300 individuals, including immunocompetent patients, immunocompromised patients, and patients with UC. Stool samples were collected from the recto-sigmoid region of the colon by endoscopy. The samples were qualitatively analyzed for bacterial and viral enteropathogens with a multiplex PCR assay using a Seeplex(®) Kit. Additional clinical and laboratory data were collected from the medical records.
RESULTS
A multiplex PCR assay detected 397 pathogens (191 bacteria and 206 viruses) in 215 samples (71.7%). The most frequently detected bacteria were Escherichia coli H7, 85 (28.3%); followed by Aeromonas spp., 43 (14.3%); and Clostridium perfringens, 36 (12.0%) samples. The most prevalent viruses were Epstein-Barr virus (EBV), 90 (30.0%); followed by human herpes virus-6 (HHV-6), 53 (17.7%); and cytomegalovirus (CMV), 37 (12.3%) samples. The prevalence rate of CMV infection was significantly higher in the immunocompromised group than in the immunocompetent group (P < 0.01). CMV infection was more common in patients with UC (26/71; 36.6%) than in the immunocompetent patients excluding UC (6/188; 3.2%) (P < 0.01). CMV infection was more prevalent in UC active patients (25/58; 43.1%) than in UC inactive patients (1/13; 7.7%) (P < 0.05). Among 4 groups which defined by the UC activity and immunosuppressive drugs, the prevalence rate of CMV infection was highest in the UC active patients with immunosuppressive drugs (19/34; 55.8%). Epstein-Barr virus (EBV) infection was more common in the immunocompromised patients excluding UC (18/41; 43.9%) than in the immunocompetent patients excluding UC (47/188; 25.0%) (P < 0.05). The simultaneous presence of CMV and EBV and/or HHV6 in UC active patients (14/58; 24.1%) was greater than in immunocompromised patients excluding UC (5/41; 12.2%) (P < 0.05).
CONCLUSION
The multiplex PCR assay that was used to analyze the stool samples in this study may serve as a non-invasive approach that can be used to exclude the possibility of CMV infection in patients with active UC who are treated with immunosuppressive therapy.
目的
评估多重聚合酶链反应（PCR）检测溃疡性结肠炎（UC）患者粪便样本中细菌和病毒肠道病原体的方法。
方法
我们前瞻性分析了300名个体，包括免疫功能正常患者、免疫功能低下患者和UC患者。通过内镜从结肠直肠乙状结肠区域采集粪便样本。使用Seeplex®试剂盒通过多重PCR检测法对样本进行细菌和病毒肠道病原体的定性分析。从病历中收集其他临床和实验室数据。
结果
多重PCR检测法在215份样本（71.7%）中检测到397种病原体（191种细菌和206种病毒）。最常检测到的细菌是大肠杆菌H7，85份样本（28.3%）；其次是气单胞菌属，43份样本（14.3%）；产气荚膜梭菌，36份样本（12.0%）。最常见的病毒是爱泼斯坦-巴尔病毒（EBV），90份样本（30.0%）；其次是人类疱疹病毒6型（HHV-6），53份样本（17.7%）；巨细胞病毒（CMV），37份样本（12.3%）。免疫功能低下组CMV感染的患病率显著高于免疫功能正常组（P<0.01）。CMV感染在UC患者中（26/71；36.6%）比在排除UC的免疫功能正常患者中（6/188；3.2%）更常见（P<0.01）。CMV感染在UC活动期患者中（25/58；43.1%）比在UC非活动期患者中（1/13；7.7%）更普遍（P<0.05）。在根据UC活动情况和免疫抑制药物定义的4组中，CMV感染的患病率在接受免疫抑制药物治疗的UC活动期患者中最高（19/34；55.8%）。爱泼斯坦-巴尔病毒（EBV）感染在排除UC的免疫功能低下患者中（18/41；43.9%）比在排除UC的免疫功能正常患者中（47/188；25.0%）更常见（P<0.05）。UC活动期患者中CMV与EBV和/或HHV6同时存在的情况（14/58；24.1%）多于排除UC的免疫功能低下患者（5/41；12.2%）（P<0.05）。
结论
本研究中用于分析粪便样本的多重PCR检测法可作为一种非侵入性方法，用于排除接受免疫抑制治疗的活动期UC患者CMV感染的可能性。
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