反转录多聚酶链反应联合间接免疫荧光法对儿童下呼吸道非典型病原体感染早期诊断的意义

Early Diagnostic Value of Reverse Transcription Polymerase Chain Reaction Combined with Indirect Immunofluorescence Method for Atypical Pathogens Infection of Lower Respiratory Tract in Children

目的评价实时反转录多聚酶链反应(RT-PCR)联合间接免疫荧光(IIF)法(联检法)对提高儿童下呼吸道非典型病原体感染检出率的效果。方法临床可疑非典型肺炎患儿152例采用联检法检测肺炎支原体(MP)、嗜肺军团菌(LP)、肺炎衣原体(CP)和Q热立克次体(QP)感染情况,同时采用间接血凝法、免疫荧光法或微量凝聚试验等多种方法进行验证。对比分析各种检测方法对MP、LP、CP和QP的检出率。并以此检验结果为依据,对所有检出的非典型病原菌感染病例使用敏感抗生素,观察治疗后患儿心率、呼吸、体温和WBC变化及临床症状改善情况。采用SPSS 13.0软件进行统计学分析。结果单独采用IIF法与RT-PCR法检测比较,对MP、LP、CP和QP的检出率差异均无统计学意义(Pa>0.05);采用联检法时,对MP和LP感染的检出率和总检出率均明显高于其余3种检测方法(Pa<0.05)。根据联检法结果选用敏感抗生素治疗,临床疗效较好。结论使用联检法可明显提高MP和LP感染的检出率和总的检出率,有效避免漏诊,但对能否提高CP和QP感染的检出率,仍有待进一步研究。

Objective To evaluate the early diagnostic value of reverse transcription polymerase chain reaction （ RT - PCR） combined with indirect immunofluorescence （IIF） method （RT - PCR ＋ IIF） for atypical pathogens infection of lower respiratory tract in children. Methods One hundred and fifty - two children with suspecting of atypical pneumonia were detected myeoplasma pneumoniae （MP） , legionella pneumophila（ LP）, ehlamydia pneumoniae （CP） and rickettsia burneti （QP）by RT- PCR ＋ IIF method;and indirect hemagglutination method, immumofluoreseence method and micro condensation experiment were performed at the same time as control. The detection ratio of MP, LP, CP and QP by every method were contrasted and analyzed, and according to these detection result, sensitive antibiotics was used for all the children with atypical pathogenic bacteria, the heart rate, respiration, body temperature and WBC changs and clinical symptom improv condi- tion of children post - treatment were observed. Results The diagnosis ratio of MP, LP, CP and QP were not different significantly between RT - PCR and IIF method（ Pa 〉 0.05 ）. While the diagnosis ratio of MP and LP with RT - PCR ＋ IIF method were higher than any of other 3 methods significantly （ P 〈 0.05 ）. According to the results by RT - PCR ＋ IIF method to use sensitive antibiotics for the children, the therapeutic efficacy was better. Conclusions RT - PCR ＋ IIF can enhance the diagnosis ratio of MP and LP, and it can well avoid the missed diagnosis,whether the diagnosis ratio of CP and QP can be enhanced need further study.