Clinical evaluation of 3 methods for the determination of Mycoplasma pneumoniae infection in children

doi:10.3969/j.issn.1673-8640.2019.09.009

Abstract

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

CLC Number:

R446.1

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.

Figures/Tables 3

References 14

[1]	JAIN S,WILLIAMS D J,ARNOLD S R,et al.Community-acquired pneumonia requiring hospitalization among U. S. children[J]. N Engl J Med,2015,372(9): 835-845.
[2]	崔明姬. 阿奇霉素在小儿肺炎支原体感染的应用效果探讨[J]. 河北医学,2014,20(1):156-158.
[3]	江载芳,申昆玲,沈颖. 诸福棠实用儿科学[M]. 8版. 北京:人民卫生出版社,2015:375-380.
[4]	TEMPLETON K E,SCHELTINGA S A,GRAFFELMAN A W,et al.Comparison and evaluation of real-time PCR,real-time nucleic acid sequence-based amplification,conventional PCR,and serology for diagnosis of Mycoplasma pneumoniae[J]. J Clin Microbiol,2003,41(9): 4366-4371.
[5]	钱月芳,唐进,陆敏. 肺炎支原体的3种实验室检查方法对照分析[J]. 临床肺科杂志,2009,14(11):1554.
[6]	MEDJO B,ATANASKOVIC-MARKOVIC M,RADIC S,et al.Mycoplasma pneumoniae as a causative agent of community-acquired pneumonia in children: clinical features and laboratory diagnosis[J]. Ital J Pediatr,2014,40:104.
[7]	KIM N H,LEE J A,EUN B W,et al.Comparison of polymerase chain reaction and the indirect particle agglutination antibody test for the diagnosis of Mycoplasma pneumoniae pneumonia in children during two outbreaks[J]. Pediatr Infect Dis J,2007,26(10): 897-903.
[8]	BARKER C E,SILLIS M,WREGHITT T G.Evaluation of Serodia Myco Ⅱ particle agglutination test for detecting Mycoplasma pneumoniae antibody:comparison with mu-capture ELISA and indirect immunofluorescence[J]. J Clin Pathol,1990,43(2): 163-165.
[9]	LOENS K,IEVEN M.Mycoplasma pneumoniae:current knowledge on nucleic acid amplification techniques and serological diagnostics[J]. Front Microbiol,2016,7: 448.
[10]	龚亚东,马庆庆,林牧. 儿童肺炎支原体感染3种检测方法对比分析[J]. 现代医药卫生,2017,33(2):251-253.
[11]	钟礼立,彭力,黄寒,等. 支气管肺泡灌洗液荧光定量PCR对儿童肺炎支原体肺炎诊断研究[J]. 中国当代儿科杂志,2011,13(3):191-194.
[12]	WANG L,FENG Z,ZHAO M,et al.A comparison study between GeXP-based multiplex-PCR and serology assay for Mycoplasma pneumoniae detection in children with community acquired pneumonia[J]. BMC Infect Dis,2017,17(1): 518.
[13]	刘剑荣,张勇,陈玲. 小儿肺炎1 224例肺炎支原体抗体检测结果分析[J]. 临床和实验医学杂志,2010,9(1):54-55.
[14]	袁艺,伏瑾,曹玲,等. 肺炎支原体荧光定量聚合酶链反应检测在儿童肺炎支原体肺炎中的诊断意义[J]. 实用儿科临床杂志,2009,24(10):760-762.

荧光定量PCR	被动凝集法		多重PCR
荧光定量PCR	+	-	+	-
+	187	3	154	36
-	22	1	8	15
χ²值	…		24.12
P值	0.369		<0.001
Kappa值	0.043		0.301

荧光定量PCR	被动凝集法		多重PCR
荧光定量PCR	+	-	+	-
+	187	3	154	36
-	22	1	8	15
χ²值	…		24.12
P值	0.369		<0.001
Kappa值	0.043		0.301

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