摘要
目的探讨PCR测序法、AS-PCR法、多重荧光PCR法检测肺炎支原体(MP)及大环内酯类耐药肺炎支原体(MRMP)的临床意义。方法采集呼吸道感染患儿的咽拭子样本116例,使用PCR测序法、AS-PCR法、多重荧光PCR法检测MP和MRMP,探讨三种方法的特点和临床价值。结果共采集116份咽拭子样本,PCR测序法检测MP阳性69例,阳性率59.48%,其中耐药株67例,包含1株耐药株与敏感株混合感染,耐药率97.1%,均为A2063G点突变,敏感株2例。AS-PCR法检测MP阳性63例,阳性率54.31%,其中耐药株58例,耐药率92.06%,敏感株5例,58例耐药株中混合感染21例,混合感染率33.33%(21/63),混合感染中18例耐药株比例超过50%。与PCR测序法比较,对MP阳性检测结果一致性极好(Kappa=0.86),对MRMP检测结果有较高的一致性(Kappa=0.652)。多重荧光PCR法检测MP阳性共67例,阳性率57.76%,其中耐药株61例,耐药率91.04%,敏感株6例,与PCR测序法相比,对MP阳性检测结果有极好的一致性(Kappa=0.929),对MRMP检测结果中度一致(Kappa=0.476)。结论PCR测序法、AS-PCR法和多重荧光PCR法检测敏感性和特异性均较高,三种方法均可同时进行MP和MRMP检测。AS-PCR法和多重荧光PCR法操作简单,适合临床应用,能为临床诊疗快速提供MP和MRMP检测结果。AS-PCR法能检测MP耐药株和敏感株混合感染,检测敏感性好。三种检测方法可为临床医生合理用药提供数据支持。
Objective We aim to compare the clinical significance and consistency of PCR sequencing, AS-PCR and multiplex fluorescence PCR in the detection of Mycoplasma pneumoniae(MP) and macrolide-resistant Mycoplasma pneumoniae(MRMP).Methods A total of 116 throat swab samples from children with respiratory tract infection were collected, and MP and MRMP were detected by PCR sequencing, AS-PCR and multiplex fluorescence PCR to evaluate their characteristics and clinical value.Results From the collected 116 samples, 69 MP-positive cases were detected by PCR sequencing method(with a positive rate of 59.48%) among which 67 drug-resistant strains were included and one mixed infection of macrolide resistant and susceptible strain was found. The drug-resistant rate was 97.1%, all of which carry A2063G point mutations. Two susceptible strains were included.63 MP-positive cases were detected by AS-PCR method(with a positive rate of 54.31%), among which 58 drug-resistant strains and 5 susceptible strains were included, with a drug-resistant rate of 92.06%. From the 58 drug-resistant strains, 21 samples were mixed infection. AS-PCR methods was consistent with PCR sequencing for detection of MP-positive samples(Kappa = 0.86) and MRMP-positive samples(Kappa = 0.652). A total of 67 MP-positive cases were detected by multiplex fluorescence PCR, with a positive rate of 57.76%, including 61 cases of drug-resistant strains and 6 cases of sensitive strains. Compared with, multiplex fluorescence PCR was highly consistent with PCR sequencing for detection of MP-positive samples(Kappa = 0.929), and moderately consistent with PCR sequencing for detection of MRMP-positive samples(Kappa = 0.476).Conclusion PCR sequencing, AS-PCR and multiplex fluorescence PCR are all highly sensitive and specific, and all the three methods can detect MP and MRMP simultaneously. AS-PCR method and multiplex fluorescence PCR method are simple and suitable for clinical application, and can provide rapid detection of MP and MRMP for clinical diagnosis and treatment. T
作者
姜越
蔚然
郑兴厂
窦海伟
涂鹏
袁青
史大伟
秦选光
辛德莉
JIANG Yue;WEI Ran;ZHENG Xing-chang;DOU Hai-wei;TU Peng;YUAN Qing;SHI Da-wei;QIN Xuan-guang;XIN De-li(Pediatric Department,Beijing Chaoyang Hospital,Capital Medical University,Beijing 100020,China;Pneumology Department,Women and Children's Hospital,Qingdao University,Shandong 266000,China;Beijing Key Laboratory for Research on Prevention and Treatment of Tropical Disease,Beijing Tropical Medicine Research Institute,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China)
出处
《中国医药生物技术》
2022年第3期226-230,共5页
Chinese Medicinal Biotechnology
基金
国家自然科学基金(81271890)。
