Detection of cytomegalovirus (CMV) by digital PCR in stool samples for the non-invasive diagnosis of CMV gastroenteritis.
Detection of cytomegalovirus (CMV) by digital PCR in stool samples for the non-invasive diagnosis of CMV gastroenteritis.
复制标题
DOI：
10.1186/s12985-022-01913-z
复制
发表时间：
2022-11-11
期刊：
影响因子：
4.8
通讯作者：
Cai, Haodong
中科院分区：
文献类型：
--
作者：
Gu, Jia;Ji, Hongyan;Liu, Tongyuan;Chen, Caixia;Zhao, Siye;Cao, Yang;Wang, Na;Xiao, Min;Chen, Liting;Cai, Haodong
CMV胃肠炎在接受异基因造血干细胞移植的患者中很常见，很难与急性移植物抗宿主病（aGvHD）区分开来，后者的症状非常相似，但需要截然不同的治疗。 CMV胃肠炎是由胃肠道局部感染或CMV重新激活引起的，而aGvHD是由于免疫排斥引起的。诊断 CMV 胃肠炎和 aGvHD 的金标准是内窥镜下胃肠道活检，这是侵入性的，可能会导致严重的副作用。使用定量聚合酶链反应（qPCR）检测粪便样本可能是一种替代方案，但在已报道的研究中，其在痕量水平测量和精度方面的应用并不都令人满意。在本研究中，我们设计了一种新方法，从粪便上清液中提取游离DNA (cfDNA)，进行数字PCR (dPCR) 检测CMV，分析其性能，并将其与当前程序提取的总DNA 进行比较。使用两种 DNA 提取方法的 22 配对粪便样本证明 cfDNA 提取方法具有明显更高的 DNA 浓度和控制基因拷贝数，这表明 cfDNA 可能提供更多信息，对于 CMV DNA 片段的检测更有用。检测 CMV DNA 片段的 dPCR 方法还表现出良好的线性度 (R2 = 0.997) 和更高的灵敏度（50% 时的检测限为 3.534 拷贝/μL）。对44名免疫功能低下患者的82份粪便样本进行分析，CMV阳性率为28%，表明这些患者中超过四分之一的胃肠道症状可能是由CMV感染或重新激活引起的。综合结果表明，通过 dPCR 检测粪便上清液 cfDNA 中的 CMV 是识别 CMV 胃肠炎的有力方法，有助于临床治疗决策。在线版本包含可在 10.1186/s12985-022-01913-z 获取的补充材料。
CMV gastroenteritis is common in patients receiving allogeneic hematopoietic stem cell transplantation and it is difficult to distinguish from acute graft-versus-host disease (aGvHD), which has very similar symptoms but needs quite different treatment. CMV gastroenteritis is caused by local infection or reactivation of CMV in the gastrointestinal tract while aGvHD is due to immune rejection. The gold standard of diagnosis of CMV gastroenteritis and aGvHD is gastrointestinal biopsy under endoscopy, which is invasive and can potentially lead to severe side effects. Stool samples testing with quantitative polymerase chain reaction (qPCR) may be an alternative, while the application in trace level measurements and precision are not all satisfactory enough in reported research. In this study, we designed a novel method that extracted the cell free DNA (cfDNA) from the fecal supernatant to perform digital PCR (dPCR) for the detection of CMV, analyzed the performance and compared it with the total DNA extracted by the current procedure. Twenty-two paired stool samples using two DNA extraction methods proved that the cfDNA extraction method had markedly higher DNA concentrations and control gene copy number, suggesting that cfDNA may be more informative and more useful for the detection of CMV DNA segment. The dPCR approach in detecting CMV DNA segment also exhibit good linearity (R2 = 0.997) and higher sensitivity (limit of detection at 50% was 3.534 copies/μL). Eighty-two stool samples from 44 immunocompromised patients were analyzed, CMV-positive rate was 28%, indicating that more than one-quarter of the gastrointestinal symptoms within these patients may be caused by CMV infection or reactivation. The combined results suggest that detection of CMV by dPCR in cfDNA of stool supernatant is a powerful method to identify CMV gastroenteritis and helps in clinical treatment decision making. The online version contains supplementary material available at 10.1186/s12985-022-01913-z.
登录
查看更多内容
影响因子：
7.7
作者：
Arora, Gaurav;Mannalithara, Ajitha;Triadafilopoulos, George
通讯作者：
Triadafilopoulos, George
DOI：
10.1016/j.diagmicrobio.2014.09.004
发表时间：
2014-12-01
影响因子：
2.9
作者：
Sedlak, Ruth Hall;Kuypers, Jane;Jerome, Keith R.
通讯作者：
Jerome, Keith R.
影响因子：
4.6
作者：
Videnska, Petra;Smerkova, Kristyna;Budinska, Eva
通讯作者：
Budinska, Eva
影响因子：
8.8
作者：
Ganzenmueller, Tina;Kluba, Jeanette;Heim, Albert
通讯作者：
Heim, Albert
影响因子：
3.1
作者：
Herfarth, Hans H.;Long, Millie D.;Miller, Melissa B.
通讯作者：
Miller, Melissa B.