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环介导等温扩增技术在检测下呼吸道感染常见病原菌中的应用

[The application of loop-mediated isothermal amplification for detection of common pathogenic bacteria in lower respiratory tract infections].

作者信息

Chen Yusheng, Wang Daxuan, Li Hongru, Chen Wenying, Zhang Xiang'e, Shao Hui, Xu Nengluan, Lin Ming, Wang Can, Yan Shenglong, Hu Xinlan, Gao Zhancheng

机构信息

The Department of Respiratpory Medicine, Fujian Provincial Hospital, Fuzhou 350001, China. Email:

Email:

出版信息

Zhonghua Jie He He Hu Xi Za Zhi. 2014 Apr;37(4):270-3.

Abstract

OBJECTIVE

To investigate the spectrum of common pathogenic bacteria of low respiratory tract infection by loop-mediated isothermal amplification (LAMP) of nucleic acid test and to prove the clinical significance of this method.

METHODS

A total of 289 qualified sputum samples from patients with lower respiratory tract infections in Fujian Province were detected by LAMP technique, and then the distribution of pathogenic bacteria was analyzed. The positive cases (the patients whose specific bacterial copies in their sputum samples > 1×10(3) copies/ml) were divided into 2 groups according to whether their treatment had covered this pathogen or not. The underlying diseases, duration of anti-bacterial treatment, the hospital days, and the effectiveness of initial treatment and cure rate were compared.

RESULTS

The culture method in the 289 patients showed that 44 (15.2%) were positive. Tests by the LAMP method with a bacteria concentration > 1×10(3) copies/ml as cutoff value, showed positive results in 124 patients (43.0%). The lower respiratory tract pathogens included 144 strains of bacteria (77.8%), and 41 strains of atypical pathogens (22.2%). Gram-negative bacteria were the predominant pathogens, such as Pseudomonas aeruginosa, H. influenzae, Klebsiella pneumoniae, and Streptococcus pneumoniae. In 95 cases the initial therapy had covered the pathogens, while in 29 cases the initial therapy had not. The effectiveness of the initial treatment (χ(2) = 31.0, P < 0.01) and the total days of hospital stay (t = -2.083, P = 0.039) in the group whose antibiotics had covered the pathogens were significantly higher than those of the other group. However, there were no significant difference in duration of anti-bacterial treatment (t = -1.073, P = 0.285)and cure rates (χ(2) = 0.6, P = 0.4) between the 2 groups.

CONCLUSIONS

LAMP method can detect the nuclear acid of the bacteria in the sputum much more rapidly and sensitively than the routine culture method. LAMP technique may be helpful to know the pathogenic bacteria before treatment, and therefore may improve the choice of initial antibiotic therapy.

摘要

目的

通过核酸环介导等温扩增(LAMP)技术研究下呼吸道感染常见病原菌谱,并验证该方法的临床意义。

方法

采用LAMP技术对福建省289例合格的下呼吸道感染患者痰液样本进行检测,分析病原菌分布情况。将阳性病例(痰液样本中特定细菌拷贝数>1×10(3)拷贝/ml的患者)根据初始治疗是否覆盖该病原菌分为两组,比较两组患者的基础疾病、抗菌治疗疗程、住院天数、初始治疗有效率及治愈率。

结果

289例患者的培养法检测结果显示,44例(15.2%)为阳性。以细菌浓度>1×10(3)拷贝/ml为临界值的LAMP法检测结果显示,124例患者(43.0%)为阳性。下呼吸道病原菌中细菌有144株(77.8%),非典型病原体有41株(22.2%)。革兰阴性菌为主要病原菌,如铜绿假单胞菌、流感嗜血杆菌、肺炎克雷伯菌和肺炎链球菌。95例患者初始治疗覆盖了病原菌,29例未覆盖。抗菌治疗覆盖病原菌组的初始治疗有效率(χ(2)=31.0,P<0.01)和总住院天数(t=-2.083,P=0.039)显著高于未覆盖组。然而,两组的抗菌治疗疗程(t=-1.073,P=0.285)和治愈率(χ(2)=0.6,P=0.4)差异无统计学意义。

结论

LAMP法比常规培养法能更快速、灵敏地检测痰液中的细菌核酸。LAMP技术有助于在治疗前了解病原菌,从而可能改善初始抗生素治疗的选择。

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