国际呼吸杂志
國際呼吸雜誌
국제호흡잡지
INTERNATIONAL JOURNAL OF RESPIRATION
2011年
8期
576-580
,共5页
细菌性肺炎%诊断%环介导等温扩增
細菌性肺炎%診斷%環介導等溫擴增
세균성폐염%진단%배개도등온확증
Bacterial pneumonia%Diagnosis%Loop-mediated isothermal amplification
目的 应用环介导等温扩增(LAMP)方法对肺炎患者痰液常见致病菌进行核酸检测,研究LAMP方法在细菌性肺炎病原学诊断中的价值.方法 抽提75例肺炎患者痰液细菌DNA,根据肺炎常见8种致病细菌设计引物,应用LAMP技术扩增DNA,实时检测样本荧光信号,对定量结果应用不同界值进行定性分析,并分别与痰培养结果进行比较.结果 LAMP技术扩增的细菌核酸产物,以细菌浓度＞1×104为界值,阳性率为68.0%,与痰培养结果符合率为56.0%;以细菌浓度＞1×105为界值,阳性率为50.7%,与痰培养结果符合率为58.7%;以细菌浓度＞1×106为界值,阳性率为30.7%,与痰培养结果符合率为53.3%.上述三组阳性率与痰培养阳性率比较,以细菌浓度＞1×105为界值组阳性率与痰培养阳性率差异无统计学意义.进一步研究在以＞1×105为界值时LAMP和痰培养方法对8种细菌的检出率,LAMP方法对部分细菌检出率高于痰培养方法,尤其对于苛养菌.结论 LAMP方法可以简便迅速扩增细菌性肺炎患者痰液中常见致病菌核酸,从而明确感染病原菌类型.LAMP扩增产物以细菌浓度＞1×105为界值,与痰培养方法比较,细菌检出率高.尤其对于苛养菌,LAMP方法有显著优势.
目的 應用環介導等溫擴增(LAMP)方法對肺炎患者痰液常見緻病菌進行覈痠檢測,研究LAMP方法在細菌性肺炎病原學診斷中的價值.方法 抽提75例肺炎患者痰液細菌DNA,根據肺炎常見8種緻病細菌設計引物,應用LAMP技術擴增DNA,實時檢測樣本熒光信號,對定量結果應用不同界值進行定性分析,併分彆與痰培養結果進行比較.結果 LAMP技術擴增的細菌覈痠產物,以細菌濃度＞1×104為界值,暘性率為68.0%,與痰培養結果符閤率為56.0%;以細菌濃度＞1×105為界值,暘性率為50.7%,與痰培養結果符閤率為58.7%;以細菌濃度＞1×106為界值,暘性率為30.7%,與痰培養結果符閤率為53.3%.上述三組暘性率與痰培養暘性率比較,以細菌濃度＞1×105為界值組暘性率與痰培養暘性率差異無統計學意義.進一步研究在以＞1×105為界值時LAMP和痰培養方法對8種細菌的檢齣率,LAMP方法對部分細菌檢齣率高于痰培養方法,尤其對于苛養菌.結論 LAMP方法可以簡便迅速擴增細菌性肺炎患者痰液中常見緻病菌覈痠,從而明確感染病原菌類型.LAMP擴增產物以細菌濃度＞1×105為界值,與痰培養方法比較,細菌檢齣率高.尤其對于苛養菌,LAMP方法有顯著優勢.
목적 응용배개도등온확증(LAMP)방법대폐염환자담액상견치병균진행핵산검측,연구LAMP방법재세균성폐염병원학진단중적개치.방법 추제75례폐염환자담액세균DNA,근거폐염상견8충치병세균설계인물,응용LAMP기술확증DNA,실시검측양본형광신호,대정량결과응용불동계치진행정성분석,병분별여담배양결과진행비교.결과 LAMP기술확증적세균핵산산물,이세균농도＞1×104위계치,양성솔위68.0%,여담배양결과부합솔위56.0%;이세균농도＞1×105위계치,양성솔위50.7%,여담배양결과부합솔위58.7%;이세균농도＞1×106위계치,양성솔위30.7%,여담배양결과부합솔위53.3%.상술삼조양성솔여담배양양성솔비교,이세균농도＞1×105위계치조양성솔여담배양양성솔차이무통계학의의.진일보연구재이＞1×105위계치시LAMP화담배양방법대8충세균적검출솔,LAMP방법대부분세균검출솔고우담배양방법,우기대우가양균.결론 LAMP방법가이간편신속확증세균성폐염환자담액중상견치병균핵산,종이명학감염병원균류형.LAMP확증산물이세균농도＞1×105위계치,여담배양방법비교,세균검출솔고.우기대우가양균,LAMP방법유현저우세.
Objective To investigate the value of loop-mediated isothermal amplification (LAMP) assay in the etiology diagnosis of bacterial pnuemonia through nucleic acid detection of common pathogens in pneumonia patients by LAMP method. Methods Sputum DNA was extracted from 75 pneumonia patients. DNA was amplified by LAMP. The fluorescence signals of products were detected by real-time PCR. The quantitative results were qualitatively analyzed at different cutoff values, and the data were compared with the results of sputum culture. Results The positive ratio of amplified products with LAMP assay at the cutoff value of 1 × 104 was 68.0% ,and the coincidence rate between LAMP assay and sputum culture was 56.0%. The positive ratio of amplified products with LAMP assay at the cutoff value of 1× 105 was 50.7%, and the coincidence rate between LAMP assay and sputum culture was 58.7%.The positive ratio of amplified products with LAMP assay at the cutoff value of 1 × 106 was 30.7%, and the coincidence rate between LAMP assay and sputum culture was 53.3%. There was no statistical significance on the positive rate between sputum culture and LAMP results at the cutoff value of 1 × 105.The detection rate of parts of bacteria by LAMP assay is higher than that by sputum culture at the cutoff value of 1 × 105 ,especially for the harsh bacteria. Conclusions DNA of common pathogens in sputum of patients with bacterial pneumonia can be easily and quickly amplified by LAMP method to identify pathogenic bacteria types. Compared with sputum culture, the bacterial detection rate is higher by LAMP assay at the cutoff value of 1 × 105. Especially for the harsh bacteria, LAMP method has significant advantages.
