巢式聚合酶链反应与病毒分离培养对呼吸道合胞病毒检测的差异及其临床意义

Difference between Nested-polymerase chain reaction and virus isolation in detection of respiratory syncytial virus and their clinical significances

目的 探讨巢式PCR（N-PCR）、病毒分离培养2种不同方法检测呼吸道合胞病毒（RSV）的差异性及其临床意义。方法 收集2010年1月至2012年8月重庆医科大学附属儿童医院呼吸病房住院患儿鼻咽抽吸物（NPAs），同时采用N-PCR、病毒分离培养方法检测 RSV。采集患儿临床资料进行统计学分析。结果 N-PCR、病毒分离培养方法同时检测RSV的标本共1143份，男女=2.161.00；年龄1～165个月（中位数为7个月）。临床主要诊断：支气管肺炎［478例（41.8%）］，间质性肺炎［223例（19.5%）］，毛细支气管炎［221例（19.3%）］，支气管炎［71例（6.2%）］，上呼吸道感染［21例（1.8%）］。N-PCR 检出 RSV 阳性共458份（总阳性率为40.1% ，其中RSV-A 31.7% ，RSV-B 7.7% ，RSV-A及RSV-B同时检出的标本占0.7%），病毒分离培养方法检出RSV 阳性标本 204份（17.8%）。分析 N-PCR 及病毒分离培养检测结果：2种方法检测均阳性（P＋I＋）标本165份，均阴性标本（P-I-）646份（一致性为70.1%），2种方法差异最大部分在于N-PCR 阳性、病毒分离培养阴性的（P＋I-）标本，共293份，占总标本数25.6% 。P＋I-组较 P-I-组患儿年龄小、住院时间长，RSV检出率冬季最高、夏季最低，发热患儿比例低，咳痰、喘息、呼吸困难、重症肺炎、呼吸衰竭患儿比例较高，差异均有统计学意义（P 均 〈0.05），临床符合 RSV 感染的特征。P＋I-组较 P＋I＋组患儿入院前病程长（P =0.005），喘息患儿比例低（P =0.009）。结论 N-PCR 和病毒分离培养2种方法对 RSV 检出的差异性与患儿入院前病程有关。N-PCR检测RSV的敏感性及特异性良好。

Objective To observe the differences between Nested-polymerase chain reaction（N-PCR） and vires isolation methods used for detection of respiratory syncytial virus （RSV） , and to reveal the potential clinical features of them. Methods From Jan. 2010 to Aug. 2012, nasopharyngeal aspirates （ NPAs ） were collected from the children with respiratory infection in the Department of Respiratory, the Children＇s Hospital of Chongqing Medical University. Both N-PCR and virus isolation were applied to detect RSV, and clinical data were collected for statistical analysis. Re- sults A total of 1143 specimens were used for RSV detection by N-PCR and virus isolation. The male-female ratio was 2.16 vs 1.00. The age of patients was ranged from 1 month to 165 months（ median：7 months）. The most common diagnoses were as follows： bronchopneumonia [ 478 cases （ 41. 8% ） ], chronic fibrous pneumonia [ 223 cases （ 19.5% ） ], bron-chiolitis [ 221 cases （ 19.3% ） ], bronchitis [ 71 cases （ 6.2% ） ] and upper respiratory infection [21 cases（ 1.8% ） ]. For N-PCR,458 cases were RSV positive （total positive rate was 40.1% ; 31.7% for RSV-A, 7.7% for RSV-B, 0. 7% for both RSV-A and RSV-B）. With virus isolation method, 204 cases were positive （ 17.8% ）. Comparison result of N-PCR and virus isolation showed： 165 cases were positive （ P ＋ I ＋ ） and 646 cases were negative （ P - I - ） by both methods （ identity was 70.1% ）, and the most difference was N-PCR positive but virus isolation negative group （ P ＋ I - ） （ 293 cases ,25.6% ）. When compared to P - I - group, the clinical features of P ＋ I - group were as follows：younger,longer hospital stays, remarkable season distribution （with peak in winter and lowest in summer） ,lower percentage of fever, higher percentage of cough, wheezing, dyspnea, severe pneumonia and respiratory failure, all these differences were statistically significant （ all P 〈 0.05 ） , the ma-nifestations matched the clinical features of RSV infection. When compared to P ＋ I＋ group, the symptoms in the P ＋ I - group had longer duration before they were admitted to hospital （ P = 0. 005 ） and lower percentage of wheezing （ P = 0. 009 ）. Conclusions The differences between N-PCR and virus isolation for the detection of RSV existed in duration of symptoms prior to hospitalization. Both the sensibility and specificity of N-PCR are desirable for RSV detection.