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非肺孢子菌肺炎患者支气管肺泡灌洗液肺孢子菌检出情况

DOI: 10.13604/j.cnki.46-1064/r.2015.10.05, PP. 1172-1176

Keywords: 耶氏肺孢子菌,非肺孢子菌肺炎,核酸扩增试验,支气管肺泡灌洗液

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Abstract:

目的利用常规PCR、巢式PCR(NestPCR,nPCR)和定量PCR(QuantitativePCR,qPCR)方法,调查非肺孢子菌肺炎(Non-Pneumocystispneumonia,non-PCP)患者支气管肺泡灌洗液(Bronchialalveolarlavagefluid,BALF)中耶氏肺孢子菌(Pneumocystisjirovecii,Pj)的检出情况。方法入选non-PCP患者50例留取BALF,分别以线粒体大亚基rRNA(MitochondriallargesubunitrRNA,mtLSUrRNA)为靶基因进行常规PCR(Mt-PCR)和巢氏PCR(Mt-nPCR),以主要表面糖蛋白(Majorsurfaceglycoprotein,Msg)为靶基因进行常规PCR(Msg-PCR),检测Mt-PCR,Mt-nPCR和Msg-PCR三种方法的Pj特异性核酸片段检出率,并对PCR检出为阳性的样本分别用以mtLSUrRNA和Msg为靶基因的qPCR来检测相应Pj核酸片段拷贝数。结果50例BALF中,Mt-nPCR、Mt-PCR及Msg-PCR的检出率分别为56%(28/50)、36%(18/50)和26%(13/50),其中三种检测方法同时阳性5例(10%),任意两种方法阳性13例(26%),任意一种方法阳性18例(36%),三种方法均为阴性14例(28%)。36例PCR检测结果阳性的标本分别进行mtLSUrRNA定量PCR(Mt-qPCR)和Msg定量PCR(Msg-qPCR)检测,其中Mt-qPCR检测结果为1000拷贝/μL~9999拷贝/μL有36例(100%);Msg-qPCR检测结果为100拷贝/μL~999拷贝/μL有3例(8.3%),1000拷贝/μL~9999拷贝/μL有16例(44.4%),10000拷贝/μL~99999拷贝/μL有9例(25%),105拷贝/μL以上8例(22.2%)。36例配对检测qPCR样本中,Msg-qPCR拷贝数大于Mt-qPCR拷贝数样本28例(77.8%)。结论在non-PCP患者BALF中,Pj核酸扩增方法的检出率为72%,Mt-qPCR拷贝数主要位于103拷贝/μL数量级,Msg-qPCR拷贝数主要分布于103至105拷贝/μL。用核酸扩增方法在BALF中检测出Pj时,临床意义解读需要谨慎。

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