LAMP十三联检在重症肺炎病原菌检测中的应用价值
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摘要
目的:探究并比较环介导恒温扩增芯片法(LAMP)在重症肺炎病原菌检测中的应用价值。方法:收集2017年1-12月本院收治的87例重症肺炎患者的分泌物标本,包括深部痰液、支气管吸出物、肺泡灌洗液,同时实施LAMP十三联检与痰培养检查,观察两种检查方法病原菌检出情况,并对比病原菌检出阳性率。结果:87例标本中痰培养检查病原菌为22例(25.29%),以铜绿假单胞菌(31.82%)为主且为单一病原菌感染;LAMP十三联检显示阳性标本41例(47.13%),其中单一病原菌感染24例(27.59%),混合病原菌感染17例(19.54%),以耐甲氧西林金黄色葡萄球菌(31.25%)及肺炎克雷伯菌(18.75%)为主,20～60岁、>60岁人群均以耐甲氧西林金黄色葡萄球菌感染为主,不同年龄段结核分枝杆菌复合群感染比较,差异有统计学意义(P<0.05);LAMP阳性率为47.13%,高于痰培养的25.29%(P<0.05),结核分枝杆菌复合群、肺炎衣原体及流感嗜血杆菌等部分病原菌痰培养未检测出。结论:在重症肺炎病原菌检测中,LAMP十三联检可为临床治疗提供可靠实验室诊断依据,患者以耐甲氧西林金黄色葡萄球菌感染为主,为提高检测准确率可根据实际情况联合其他检测方法,指导用药。
Objective:To explore and compare the application value of loop-mediated isothermal amplification(LAMP)in pathogenic bacteria detection of severe pneumonia.Method:Secretion specimens of87 patients with severe pneumonia admitted to our hospital from January 2017 to December 2017 were collected,it included deep sputum,bronchial aspirates and alveolar lavage fluid,at the same time,LAMP and sputum culture were carried out,the detection of pathogenic bacteria by two methods were observed,and the positive rate of pathogenic bacteria was compared.Result:Of 87 specimens,22 cases(25.29%)were pathogenic bacteria by sputum culture,pseudomonas aeruginosa(31.82%)was the main pathogen and only one pathogen was infected.LAMP showed that 41 cases(47.13%)positive specimens,including 24 cases(27.59%)of single pathogen infection,17 cases(19.54%)of mixed pathogen infection,mainly methicillin-resistant Staphylococcus aureus(31.25%)and Klebsiella pneumoniae(18.75%).People aged 20-60 years and over 60 years were mainly methicillin-resistant Staphylococcus aureus infection,with tuberculosis branches in different age groups,there was significant difference in Bacillus complex infection(P<0.05),the positive rate of LAMP in lower was 47.13% higher than 25.29% in sputum culture(P<0.05),some pathogenic bacteria such as Mycobacterium tuberculosis complex,Chlamydia pneumoniae and Haemophilus influenzae were not detected in sputum culture.Conclusion:In the detection of pathogenic bacteria in severe pneumonia,LAMP can provide reliable laboratory diagnostic basis for clinical treatment,methicillin-resistant Staphylococcus aureus infection is the main pathogen in patients.In order to improve the detection accuracy,other detection methods can be combined according to the actual situation to guide drug use.
Objective:To explore and compare the application value of loop-mediated isothermal amplification(LAMP)in pathogenic bacteria detection of severe pneumonia.Method:Secretion specimens of87 patients with severe pneumonia admitted to our hospital from January 2017 to December 2017 were collected,it included deep sputum,bronchial aspirates and alveolar lavage fluid,at the same time,LAMP and sputum culture were carried out,the detection of pathogenic bacteria by two methods were observed,and the positive rate of pathogenic bacteria was compared.Result:Of 87 specimens,22 cases(25.29%)were pathogenic bacteria by sputum culture,pseudomonas aeruginosa(31.82%)was the main pathogen and only one pathogen was infected.LAMP showed that 41 cases(47.13%)positive specimens,including 24 cases(27.59%)of single pathogen infection,17 cases(19.54%)of mixed pathogen infection,mainly methicillin-resistant Staphylococcus aureus(31.25%)and Klebsiella pneumoniae(18.75%).People aged 20-60 years and over 60 years were mainly methicillin-resistant Staphylococcus aureus infection,with tuberculosis branches in different age groups,there was significant difference in Bacillus complex infection(P<0.05),the positive rate of LAMP in lower was 47.13% higher than 25.29% in sputum culture(P<0.05),some pathogenic bacteria such as Mycobacterium tuberculosis complex,Chlamydia pneumoniae and Haemophilus influenzae were not detected in sputum culture.Conclusion:In the detection of pathogenic bacteria in severe pneumonia,LAMP can provide reliable laboratory diagnostic basis for clinical treatment,methicillin-resistant Staphylococcus aureus infection is the main pathogen in patients.In order to improve the detection accuracy,other detection methods can be combined according to the actual situation to guide drug use.
引文
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[10]李永红.痰培养标本不合格原因分析及对策[J].现代临床护理,2014,13(5):21-23.
[11]陈愉生,王大璇,李鸿茹,等.环介导等温扩增技术在下呼吸道感染常见病原体检测中的应用[J].中华结核和呼吸杂志,2014,37(4):270-273.
[12]Yasuharabell J,Silva A D,Heuchelin S A,et al.Detection of Goss’s Wilt Pathogen Clavibacter michiganensis subsp.nebraskensis in Maize by Loop-Mediated Amplification[J].Phytopathology,2015,106(3):226.
[13]吴丹丹,李保胜,亢继文,等.RT-LAMP与LAMP法检测结核分枝杆菌的效果比较[J].中国人兽共患病学报,2018,34(3):248-254.
[14]Yasuhara-Bell J,Baysal-Gurel F,Miller S A,et al.Utility of a loop-mediated amplification assay for detection of Clavibacter michiganensis subsp.michiganensis in seeds and plant tissues[J].Canadian Journal of Plant Pathology,2015,37(3):260-266.
[15]唐睿珠,罗正琼,徐秋月,等.呼吸道病原体核酸恒温扩增芯片十三联检在下呼吸道感染常见病原体基因中的检测[J].昆明医科大学学报,2017,38(1):8-12.
[2]王文欣,周茄,张宇,等.EICU重症肺炎患者病原学与耐药性分析[J].中华医院感染学杂志,2017,27(17):3845-3847.
[3]王亚军,徐红,李桂香.下呼吸道感染患者痰培养病原菌分布与耐药分析[J].解放军预防医学杂志,2017,35(6):706.
[4]Kubota R,Jenkins D M.Real-Time Duplex Applications of LoopMediated AMPlification(LAMP)by Assimilating Probes[J].International Journal of Molecular Sciences,2015,16(3):4786-4799.
[5]Charles P G P,Davis J S,Grayson M L.Rocket Science and the Infectious Diseases Society of America/American Thoracic Society(IDSA/ATS)Guidelines for Severe Community-Acquired Pneumonia[J].Clinical Infectious Diseases,2009,48(12):1796-1796.
[6]尚红,王毓三,申子瑜.全国临床检验操作规程[M].北京:人民卫生出版社,2015:123-154.
[7]郭霞,喻昌利,安庆丽,等.老年重症肺炎患者病原学分布及预后危险因素分析[J].广东医学,2016,37(6):873-875.
[8]Tomczyk S,Jain S,Bramley A M,et al.Antibiotic Prescribing for Adults Hospitalized in the Etiology of Pneumonia in the Community Study[J].Open Forum Infectious Diseases,2017,4(2):609-620.
[9]张曼,徐艳,杨怀,等.医院感染病原学诊断水平现状分析与展望[J].中华医院感染学杂志,2017,27(8):1725-1728.
[10]李永红.痰培养标本不合格原因分析及对策[J].现代临床护理,2014,13(5):21-23.
[11]陈愉生,王大璇,李鸿茹,等.环介导等温扩增技术在下呼吸道感染常见病原体检测中的应用[J].中华结核和呼吸杂志,2014,37(4):270-273.
[12]Yasuharabell J,Silva A D,Heuchelin S A,et al.Detection of Goss’s Wilt Pathogen Clavibacter michiganensis subsp.nebraskensis in Maize by Loop-Mediated Amplification[J].Phytopathology,2015,106(3):226.
[13]吴丹丹,李保胜,亢继文,等.RT-LAMP与LAMP法检测结核分枝杆菌的效果比较[J].中国人兽共患病学报,2018,34(3):248-254.
[14]Yasuhara-Bell J,Baysal-Gurel F,Miller S A,et al.Utility of a loop-mediated amplification assay for detection of Clavibacter michiganensis subsp.michiganensis in seeds and plant tissues[J].Canadian Journal of Plant Pathology,2015,37(3):260-266.
[15]唐睿珠,罗正琼,徐秋月,等.呼吸道病原体核酸恒温扩增芯片十三联检在下呼吸道感染常见病原体基因中的检测[J].昆明医科大学学报,2017,38(1):8-12.