实时荧光定量PCR法对ICU患者痰液标本中鲍曼不动杆菌耐药基因的检测及其评价

The Quantitative Real-time PCR on the Detection and Assessment of Acinetobacter Baumanii Drug Resistance Genes in The Sputum Samples of ICU Patients

目的研究实时荧光定量聚合酶链式反应(PCR)法在重症监护室(ICU)患者痰液标本中检测鲍曼不动杆菌耐药基因的效果。方法选择2021年3月至2022年12月南阳市中心医院纳入的68例ICU患者进入试验,分别收集其痰液标本,通过实时荧光定量PCR法测定标本内鲍曼不动杆菌耐药基因情况,统计鲍曼不动杆菌及耐碳青霉烯类鲍曼不动杆菌的检出率,并观察耐碳青霉烯类鲍曼不动杆菌的药敏试验结果,最后分析OXA-51基因检查结果和耐药基因OXA-23检查结果。结果68例ICU患者的痰液标本中,通过传统培养方式检出鲍曼不动杆菌33株,阳性检出率48.53%;耐碳青霉烯类鲍曼不动杆菌共检出23株,阳性检出率33.82%;基因检测OXA-51阳性显示鲍曼不动杆菌有37株,阳性检出率54.41%;OXA-23阳性显示耐碳青霉烯类鲍曼不动杆菌有25株,阳性检出率36.76%。针对碳青霉烯类药物产生耐药的鲍曼不动杆菌所占比例占75.76%。耐碳青霉烯类鲍曼不动杆菌对于大部分药物耐药,耐药性较低的药物分别有头孢哌酮舒巴坦、米诺环素、替加环素、黏菌素等。传统培养和耐药基因的阳性检出率相比,差异并无统计意义(P>0.05);经Kappa检验显示为0.879,证实两种方式的检查结果的一致性较好。传统培养和耐药基因的阳性检出率相比,差异并无统计意义(P>0.05);经Kappa检验显示为0.712,证实两种方式的检查结果的一致性一般。结论实时荧光定量PCR法的效果明显,可成为鲍曼不动杆菌耐药基因检测的主要方式。

Objective To study the effect of the quantitative real-time polymerase chain reaction(PCR)on the detection and assessment of acinetobacter baumanii drug resistance genes in the sputum samples of intensive care unit(ICU)patients.Methods A total of 68 ICU patients enrolled in Nanyang Central Hospital from March 2021 to December 2022 were selected for the trial,and sputum samples were collected respectively.The quantitative real-time PCR was applied to detect the acinetobacter baumanii drug resistance genes.The detection rates of acinetobacter baumanii and carbapenem-resistant acinetobacter baumanii were counted;the drug sensitive test results of carbapenem-resistant acinetobacter baumanii were observed.OXA-15 genes and OXA-23 drug-resistant genes were analyzed.Results In sputum samples of 68 ICU patients,33 strains of Acinetobacter baumannii were detected by traditional culture,and the positive detection rate was 48.53%.A total of 23 carbapenem-resistant Acinetobacter baumannii were detected,and the positive detection rate was 33.82%.OXA-51 gene test showed 37 strains of Acinetobacter baumannii(54.41%).OXA-23 positive results showed that there were 25 carbapenem-resistant Acinetobacter baumannii,and the positive detection rate was 36.76%.The proportion of Acinetobacter baumannii resistant to carbapenems accounted for 75.76%.Carbapenem-resistant Acinetobacter baumannii was resistant to most drugs,and the drugs with low resistance were cefoperazone sulbactam,minocycline,tigacycline and colistin.There were no significant difference about the positive bacterial detection rates based on the conventional cultivation and OXA-51 drug-resistance genes(P>0.05);Kappa values were 0.879,demonstrating the good consistency based on two examination modes.There were no significant difference about the positive bacterial detection rates based on the conventional cultivation and OXA-23 drug-resistance genes(P>0.05);Kappa values were 0.712,demonstrating the general consistency based on two examination modes.Conclusion The quantitative real-time PCR method has obvious effect,and can be the main method to detect the drug resistance gene of Acinetobacter baumannii.