百日咳临床流行病学特点及诊断方法的比较研究
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摘要
目的分析临床和实验室诊断百日咳病例的临床流行病学特点,并比较细菌培养与荧光定量PCR 2种病原菌检测方法的应用价值。方法收集北京儿童医院864例百日咳送检患儿的临床信息,并采用细菌培养和荧光定量PCR检测其鼻咽拭子标本,分析不同诊断标准下百日咳临床流行病学特点,以及2种实验室诊断的临床应用价值。结果临床诊断与细菌培养和荧光定量PCR实验室诊断的百日咳病例的地区、季节和年龄分布相似,北京儿童医院就诊的百日咳患儿以河北(41.6%～45.2%)、北京(36.4%～42.9%)两地为主,诊断病例集中于夏秋季节(5月份-9月份),婴儿是百日咳发病的主要人群(77.5%～89.3%)。2种实验室诊断方法相比,典型临床症状"吸气性回声"在细菌培养阳性样本组更为常见(X~2=5.086,P=0.024)。以临床诊断为标准,细菌培养和荧光定量PCR诊断百日咳的灵敏度分别为27.1%和51.5%,特异度分别为79.0%和59.0%;不满足临床诊断标准的458例患儿中,细菌培养或荧光定量PCR检测为阳性的共193例(42.1%)。结论临床诊断和实验室诊断百日咳揭示的临床流行病学特征相似。以临床诊断为标准,细菌培养诊断百日咳灵敏度低、特异度高,荧光定量PCR灵敏度较高、特异度较低。实验室检测能发现更多临床不典型病例。
Objective To analyze the clinical epidemiological characteristics of clinical and laboratory diagnosis of pertussis cases,and to compare the application value of two kinds of pathogen detection methods:bacterial culture and quantitative real-time PCR.(RT-qPCR). Methods The clinical information of 864 children with pertussis in Beijing Children s Hospital was collected. The nasopharyngeal swab specimens were detected by bacterial culture and RT-qPCR. The clinical epidemiological characteristics of pertussis under different diagnostic criteria and two laboratory diagnoses were analyzed. Results The regional,seasonal and age distributions of pertussis cases diagnosed by laboratory diagnosis and bacterial culture and RT-qPCR were similar. The children with whooping cough in Beijing Children s Hospital are mainly in Hebei(41.6% ～45.2%) and Beijing(36.4% ～42.9%). The diagnosed cases are concentrated in summer and autumn(May-September),and the baby is the main population of whooping cough(77.5%～89.3%). Compared to the two laboratory diagnostic methods,the typical clinical symptom"inhalation echo"is more common in the bacterial culture positive sample group(X~2=5.086, P=0.024). Based on clinical diagnosis,the sensitivity of bacterial culture and RT-qPCR for the diagnosis of whooping cough was 27.1% and 51.5%,and the specificity was 79.0% and 59.0% respectively. Among 458 children who did not meet the clinical diagnostic criteria,193(42.1%) were positive by bacterial culture or RT-qPCR. Conclusion Clinical diagnostic criteria and laboratory diagnostic criteria of pertussis revealed the similar epidemiological characteristics. Bacterial culture has a high specificity and low sensitivity,but RT-qPCR has a high sensitivity and low specificity. Laboratory detection can find more atypical pertussis cases.
Objective To analyze the clinical epidemiological characteristics of clinical and laboratory diagnosis of pertussis cases,and to compare the application value of two kinds of pathogen detection methods:bacterial culture and quantitative real-time PCR.(RT-qPCR). Methods The clinical information of 864 children with pertussis in Beijing Children s Hospital was collected. The nasopharyngeal swab specimens were detected by bacterial culture and RT-qPCR. The clinical epidemiological characteristics of pertussis under different diagnostic criteria and two laboratory diagnoses were analyzed. Results The regional,seasonal and age distributions of pertussis cases diagnosed by laboratory diagnosis and bacterial culture and RT-qPCR were similar. The children with whooping cough in Beijing Children s Hospital are mainly in Hebei(41.6% ～45.2%) and Beijing(36.4% ～42.9%). The diagnosed cases are concentrated in summer and autumn(May-September),and the baby is the main population of whooping cough(77.5%～89.3%). Compared to the two laboratory diagnostic methods,the typical clinical symptom"inhalation echo"is more common in the bacterial culture positive sample group(X~2=5.086, P=0.024). Based on clinical diagnosis,the sensitivity of bacterial culture and RT-qPCR for the diagnosis of whooping cough was 27.1% and 51.5%,and the specificity was 79.0% and 59.0% respectively. Among 458 children who did not meet the clinical diagnostic criteria,193(42.1%) were positive by bacterial culture or RT-qPCR. Conclusion Clinical diagnostic criteria and laboratory diagnostic criteria of pertussis revealed the similar epidemiological characteristics. Bacterial culture has a high specificity and low sensitivity,but RT-qPCR has a high sensitivity and low specificity. Laboratory detection can find more atypical pertussis cases.
引文
[1]姚开虎,邓继岿,热夏提·达吾提.百日咳诊断:现行标准的局限性和GPI建议[J].中国当代儿科杂志,2016,18(9):891-896.
[2]Bruno A,Almeida M,Ruiz G.A place for Bordetella pertussis in PCR-based diagnosis of community-acquired pneumonia[J].Infect Dis(Lond),2018,50(3):232-235.
[3]Mohammad R,Rusung T,Ghada N,et al.Evaluation of amplification targets for the specific detection of Bordetella pertussis using real-time polymerase chain reaction[J].Can J Infect Dis Med Microbiol,2014,25(4):217-221.
[4]Centers for Disease Control and Prevention(CDC).Manual for the surveillance of vaccine-preventable diseases:chapter 10:pertussis[EB/OL].http://www.cdc.gov/vaccines/pubs/surv-manual/chpt10-pertussis.html#laboratory,2014-01-01/2019-05-24.
[5]Centers for Disease Control and Prevention(CDC).Proper Technique for obtaining a nasopharyngeal specimen for isolation of Bordetella pertussis[EB/OL].https://www.cdc.gov/pertussis/clinical/diagnostic-testing/specimen-collection.html#1,2017-08-07/2019-05-24.
[6]宁桂军,高源,吴丹,等.中国2011-2017年百日咳流行病学特征分析[J].中国疫苗和免疫,2018,24(3):264-273.
[7]Lee A,Cassiday P,Pawloski L,et al.Clinical evaluation and validation of laboratory methods for the diagnosis of Bordetella pertussis infection:Culture,polymerase chain reaction(PCR)and anti-pertussis toxin IgG serology(IgG-PT)[J].Plos One,2018,13(4):e0195979.
[8]Schmidt G,Liese J,Porter F,et al.Polymerase chain reaction(PCR)compared with conventional identification in culture for detection of Bordetella pertussis in7153 children[J].Clinical Microbiology&Infection,2010,3(4):462-467.
[9]袁林,俞桑洁,姚开虎,等.聚合酶链反应和细菌培养及血清学方法诊断百日咳的比较研究[J].中国疫苗和免疫,2012,18(4):354-357.
[10]王增国,杨杨,刘莹,等.百日咳实验室诊断方法的应用分析与比较[J].中华流行病学杂志,2013,34(10):1010-1012.
[11]邓继岿,王红梅,田树凤.儿童百日咳的临床特点及实验室诊断[J].中华实用儿科临床杂志,2017,32(22):1692-1695.
[12]Martini H,Detemmerman L,Soetens O,et al.Improving specificity of Bordetella pertussis detection using a four target real-time PCR[J].Plos One,2017,12(4):e0175587.
[13]李亚绒.百日咳治疗研究进展[J].中华实用儿科临床杂志,2017,32(22):1696-1698.
[14]Bechini A,Tiscione E,Boccalini S,et al.Acellular pertussis vaccine use in risk groups(adolescents,pregnant women,newborns and health care workers):a review of evidences and recommendations[J].Vaccine,2012,30(35):5179-5190.
[15]World Health Organization(WHO).WHO-recommended standards for surveillance of selected vaccinepreventable diseases[EB/OL].https://www.who.int/immunization/monitoring_surveillance/burden/vpd/WHO_SurveillanceVaccinePreventable_16_Pertussis_R2.pdf ua=1,2018-09-05/2019-05-24.
[2]Bruno A,Almeida M,Ruiz G.A place for Bordetella pertussis in PCR-based diagnosis of community-acquired pneumonia[J].Infect Dis(Lond),2018,50(3):232-235.
[3]Mohammad R,Rusung T,Ghada N,et al.Evaluation of amplification targets for the specific detection of Bordetella pertussis using real-time polymerase chain reaction[J].Can J Infect Dis Med Microbiol,2014,25(4):217-221.
[4]Centers for Disease Control and Prevention(CDC).Manual for the surveillance of vaccine-preventable diseases:chapter 10:pertussis[EB/OL].http://www.cdc.gov/vaccines/pubs/surv-manual/chpt10-pertussis.html#laboratory,2014-01-01/2019-05-24.
[5]Centers for Disease Control and Prevention(CDC).Proper Technique for obtaining a nasopharyngeal specimen for isolation of Bordetella pertussis[EB/OL].https://www.cdc.gov/pertussis/clinical/diagnostic-testing/specimen-collection.html#1,2017-08-07/2019-05-24.
[6]宁桂军,高源,吴丹,等.中国2011-2017年百日咳流行病学特征分析[J].中国疫苗和免疫,2018,24(3):264-273.
[7]Lee A,Cassiday P,Pawloski L,et al.Clinical evaluation and validation of laboratory methods for the diagnosis of Bordetella pertussis infection:Culture,polymerase chain reaction(PCR)and anti-pertussis toxin IgG serology(IgG-PT)[J].Plos One,2018,13(4):e0195979.
[8]Schmidt G,Liese J,Porter F,et al.Polymerase chain reaction(PCR)compared with conventional identification in culture for detection of Bordetella pertussis in7153 children[J].Clinical Microbiology&Infection,2010,3(4):462-467.
[9]袁林,俞桑洁,姚开虎,等.聚合酶链反应和细菌培养及血清学方法诊断百日咳的比较研究[J].中国疫苗和免疫,2012,18(4):354-357.
[10]王增国,杨杨,刘莹,等.百日咳实验室诊断方法的应用分析与比较[J].中华流行病学杂志,2013,34(10):1010-1012.
[11]邓继岿,王红梅,田树凤.儿童百日咳的临床特点及实验室诊断[J].中华实用儿科临床杂志,2017,32(22):1692-1695.
[12]Martini H,Detemmerman L,Soetens O,et al.Improving specificity of Bordetella pertussis detection using a four target real-time PCR[J].Plos One,2017,12(4):e0175587.
[13]李亚绒.百日咳治疗研究进展[J].中华实用儿科临床杂志,2017,32(22):1696-1698.
[14]Bechini A,Tiscione E,Boccalini S,et al.Acellular pertussis vaccine use in risk groups(adolescents,pregnant women,newborns and health care workers):a review of evidences and recommendations[J].Vaccine,2012,30(35):5179-5190.
[15]World Health Organization(WHO).WHO-recommended standards for surveillance of selected vaccinepreventable diseases[EB/OL].https://www.who.int/immunization/monitoring_surveillance/burden/vpd/WHO_SurveillanceVaccinePreventable_16_Pertussis_R2.pdf ua=1,2018-09-05/2019-05-24.
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