LAMP-CRISPR/Cas12a技术在重症肺炎病原体检测中的应用  

Application of LAMP-CRISPR/Cas12a technology in pathogen detection for severe pneumonia

作　　者：李玲玲 匡歆 王贵平 于风格 LI Lingling;KUANG Xin;WANG Guiping;YU Fengge(Department of Clinical Laboratory,Jiaozhou Central Hospital of Qingdao,Qingdao,Shandong 266399,China;Emergency Center of Qingdao Jimo District,Qingdao,Shandong 266299,China)

机构地区：[1]山东省青岛市胶州中心医院检验科,山东青岛266399 [2]山东省青岛市即墨区急救中心,山东青岛266299

出　　处：《检验医学与临床》2025年第3期415-419,423,共6页Laboratory Medicine and Clinic

基　　金：山东省青岛市医药卫生科研指导项目(2022-WJZD124)。

摘　　要：目的分析环介导恒温扩增技术(LAMP)联合CRISPR/Cas12a技术在的重症肺炎病原学检测中的应用效果。方法选取2019年1月至2023年6月青岛市胶州中心医院收治的147例重症肺炎患者作为研究对象,取患者支气管肺泡灌洗液或气管内抽吸物标本进行临床病原学检测、血清学检测及LAMP-CRISPR/Cas12a检测,观察3种方法检测结果,比较LAMP-CRISPR/Cas12a检测方法与临床病原诊断结果、血清学检查结果一致性。结果147例患者共获得呼吸道标本147份,其中103份为泡灌洗液标本,44份为呼吸道深部痰液。临床病原学检测病原体标本100例,阳性率为68.03%。临床病原学检测结果具体为单一病原体感染85例,混合感染15例;未检测出病原体47例。147例患者中检测免疫球蛋白M抗体,共检出78例,阳性率为53.06%,其中细菌0例,非典型病原体32例,单一病毒感染32例,混合感染例14例,未检测出病原体69例。LAMP-CRISPR/Cas12a检测技术可快速检测重症肺炎病原体。LAMP-CRISPR/Cas12a检测中共检测出病原体标本117例,阳性率为79.59%,其中单纯细菌感染22例,非典型病原体26例,单纯病毒感染51例,混合感染18例,未检测出病原体30例。3种检测方法中,LAMP-CRISPR/Cas12a检测的阳性率最高,血清学检测的结果最低。且LAMP-CRISPR/Cas12a检测的阳性率(79.59%)高于临床病原学检测(68.03%)、血清学检测(53.06%),差异均有统计学意义(χ^(2)=5.085、23.164,P=0.024、<0.001)。在重症肺炎病原体检测中,LAMP-CRISPR/Cas12a检测方法与临床病原诊断结果、血清学检查结果一致性均较高(Kappa均>0.75,P<0.05)。结论LAMP-CRISPR/Cas12a检测方法在重症肺炎病原学诊断中的阳性率较高,尤其适用于细菌性重症肺炎与病毒性重症肺炎检测。Objective To analyze the application effect of loop-mediated isothermal amplification(LAMP)combined with CRISPR/Cas12a technology in the etiological detection of severe pneumonia.Methods A total of 147 patients with severe pneumonia admitted to Jiaozhou Central Hospital of Qingdao from January 2019 to June 2023 were selected as the research objects.Bronchoalveolar lavage fluid or endotracheal aspirates of the patients were collected for clinical pathogen detection,serological detection and LAMP-CRISPR/Cas12a detection,and the results of the three methods were observed.The consistency of LAMP-CRISPR/Cas12a detection method with clinical pathogen diagnosis and serological test results was compared.Results A total of 147 respiratory specimens were obtained from 147 patients,including 103 bubble lavage fluid samples and 44 deep respiratory sputum specimens.The positive rate of clinical pathogen detection was 68.03%.The clinical pathogen detection results were single pathogen infection in 85 cases and mixed infection in 15 cases.No pathogen was detected in 47 cases.Immunoglobulin M antibody was detected in 78 of 147 patients,with a positive rate of 53.06%,including 0 case of bacteria,32 cases of atypical pathogens,32 cases of single virus infection,14 cases of mixed infection,and 69 cases of no pathogen was detected.LAMP-CRISPR/Cas12a assay can rapidly detect pathogens of severe pneumonia.A total of 117 cases of pathogens were detected by LAMP-CRISPR/Cas12a assay,with a positive rate of 79.59%,including 22 cases of simple bacterial infection,26 cases of atypical pathogens,51 cases of simple viral infection,18 cases of mixed infection,and 30 cases of no pathogens were detected.LAMP-CRISPR/Cas12a assay showed the highest positive rate and serological assay showed the lowest results among the three assays.The positive rate of LAMP-CRISPR/Cas12a detection(79.59%)was higher than that of clinical etiological detection(68.03%)and serological detection(53.06%),and the differences were statistically significant(χ^(2)=5.085,23

关 键 词：重症肺炎 环介导恒温扩增技术 CRISPR/Cas12a技术 病原学 

分 类 号：R446.5[医药卫生—诊断学] R373.1[医药卫生—临床医学]