3种方法检测儿童肺炎支原体感染的临床评估

doi:10.3969/j.issn.1673-8640.2019.09.009

摘要/Abstract

摘要：

目的比较3种检测肺炎支原体（MP）感染的方法[被动凝集法、多重聚合酶链反应（PCR）及荧光定量PCR]的差异,探讨不同方法在不同年龄段及不同病程肺炎患儿中的应用价值。方法选取接受纤维支气管镜灌洗治疗的MP肺炎患儿213例。采用被动凝集法、多重PCR及荧光定量PCR分别检测血清MP抗体、痰液MP DNA及肺泡灌洗液（BALF）中的MP DNA。以荧光定量PCR在BALF中检出MP DNA为病原学判断标准,比较3种方法的一致性。将所有患儿按年龄分为<1岁组（8例）、1~3岁组（29例）、4~6岁组（97例）、>6岁组（79例）,按病程分为≤7 d组和>7 d组。比较各组3种方法的阳性检出率。结果 213例MP肺炎患儿中,被动凝集法、多重PCR及荧光定量PCR的阳性检出率分别为98.12%、76.06%和89.20%。以荧光定量PCR在BALF中检出MP DNA为病原学判断标准,多重PCR与荧光定量PCR的Kappa值为0.301,优于被动凝集法与荧光定量PCR的Kappa值（0.043）。按年龄分组,3种方法的阳性检出率均与年龄有关（P<0.05）。病程>7 d组荧光定量PCR阳性率高于≤7 d组（P=0.003）,而被动凝集法和多重PCR的阳性检出率2个组之间差异无统计学意义（P>0.05）。结论在使用纤维支气管镜灌洗治疗的MP肺炎患儿中,荧光定量PCR与多重PCR的一致性优于被动凝集法,荧光定量PCR的阳性检出率与患儿病程相关。

关键词: 肺炎支原体, 多重聚合酶链反应, 荧光定量聚合酶链反应, 肺泡灌洗液

Abstract:

Objective To compare the differences among 3 methods [passive agglutination,multiplex polymerase chain reaction（PCR） and real-time quantitation PCR] for the determination of Mycoplasma pneumoniae（MP） infection,and to investigate their roles in different ages and disease stages of children with pneumonia. Methods A total of 213 children with MP pneumonia treated with fiberoptic bronchoscopy were enrolled. The passive agglutination,multiplex PCR and real-time quantitation PCR were used to determine serum MP antibodies,MP DNA in sputum and MP DNA in the bronchoalveolar lavage fluid（BALF）,respectively. The results of real-time quantitation PCR for MP DNA in BALF were regarded as gold standard. The consistency of the 3 methods was compared. All the children were classified into <1-year-old group（8 cases）,1-3-year-old group（29 cases）,4-6-year-old group（97 cases） and >6-year-old group（79 cases）. According to disease courses,they were classified into ≤7 d and >7 d groups. The positive determination rates of the 3 methods were compared. Results Among the 213 children with MP pneumonia,the positive determination rates of passive agglutination,multiplex PCR and real-time quantitation PCR were 98.12%,76.06% and 89.20%,respectively. The results of real-time quantitation PCR for MP DNA in BALF were regarded as gold standard. The Kappa value of multiplex PCR with real-time quantitation PCR（0.301） was higher than that of passive agglutination with real-time quantitation PCR（0.043）. The positive determination rates of the 3 methods were related to ages（P<0.05）. The positive determination rate of real-time quantitation PCR in >7 d group was higher than that in ≤7 d group（P=0.003）,while the positive determination rates of passive agglutination and multiplex PCR had no statistical significance（P>0.05）. Conclusions Among MP pneumonia patients treated with fiberoptic bronchoscopy,the consistency of real-time quantitation PCR and multiplex PCR is superior to passive agglutination,and the positive determination rate of real-time quantitation PCR is related with disease courses.

Key words: Mycoplasma pneumoniae, Multiplex polymerase chain reaction, Real-time quantitation polymerase chain reaction, Bronchoalveolar lavage fluid

中图分类号:

R446.1

冯志山, 李贵霞, 刘建华, 郭巍巍, 张文超, 张丽霞, 黄慧, 王乐. 3种方法检测儿童肺炎支原体感染的临床评估[J]. 检验医学, 2019, 34(9): 808-811.

FENG Zhishan, LI Guixia, LIU Jianhua, GUO Weiwei, ZHANG Wenchao, ZHANG Lixia, HUANG Hui, WANG Le. Clinical evaluation of 3 methods for the determination of Mycoplasma pneumoniae infection in children[J]. Laboratory Medicine, 2019, 34(9): 808-811.

图/表 3

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组别	例数	被动凝集法		多重PCR		荧光定量PCR
组别	例数	阳性例数	百分比（%）	阳性例数	百分比（%）	阳性例数	百分比（%）
<1岁组	8	6	75	6	75	6	75
1~3岁组	29	29	100^*	18	62	22	76
4~6岁组	97	96	99^*	70	72	86	89
>6岁组	79	78	99^*	68	86^#	76	96^#
合计	213	209	98	162	76	190	89
χ²值		24.304		8.282		11.082
P值		<0.001		0.041		0.011

组别	例数	被动凝集法		多重PCR		荧光定量PCR
组别	例数	阳性例数	百分比（%）	阳性例数	百分比（%）	阳性例数	百分比（%）
<1岁组	8	6	75	6	75	6	75
1~3岁组	29	29	100^*	18	62	22	76
4~6岁组	97	96	99^*	70	72	86	89
>6岁组	79	78	99^*	68	86^#	76	96^#
合计	213	209	98	162	76	190	89
χ²值		24.304		8.282		11.082
P值		<0.001		0.041		0.011

组别	例数	被动凝集法		多重PCR		荧光定量PCR
		阳性例数	百分比（%）	阳性例数	百分比（%）	阳性例数	百分比（%）
≤7 d组	113	109	96	82	68	94	83
>7 d组	100	100	100	80	86	96	96
χ²值		1.942		1.610		9.044
P值		0.163		0.205		0.003

组别	例数	被动凝集法		多重PCR		荧光定量PCR
		阳性例数	百分比（%）	阳性例数	百分比（%）	阳性例数	百分比（%）
≤7 d组	113	109	96	82	68	94	83
>7 d组	100	100	100	80	86	96	96
χ²值		1.942		1.610		9.044
P值		0.163		0.205		0.003

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