中华生物医学工程杂志
中華生物醫學工程雜誌
중화생물의학공정잡지
CHINESE JOURNAL OF BIOMEDICAL ENGINEERING
2007年
6期
349-352
,共4页
杨淳%黄红川%李冰%刘启才%彭燕%袁锦屏%杨灵%苏丹红%邱贵霞%杨其霖%黎毅敏
楊淳%黃紅川%李冰%劉啟纔%彭燕%袁錦屏%楊靈%囌丹紅%邱貴霞%楊其霖%黎毅敏
양순%황홍천%리빙%류계재%팽연%원금병%양령%소단홍%구귀하%양기림%려의민
克柔念珠菌%念珠菌,光滑%聚含酶链反应
剋柔唸珠菌%唸珠菌,光滑%聚含酶鏈反應
극유념주균%념주균,광활%취함매련반응
Candida krusei%Candida glabruta%Polymerase chain reaction
目的 建立一种快速、灵敏、特异的鉴定克柔念珠菌和光滑念珠菌的双重实时荧光定量PCR方法.方法 以核糖体基因内转录间隔区Ⅱ(ITSⅡ)为靶目标,设计并合成分别针对克柔念珠菌、光滑念珠菌的种特异引物和探针.建立双重实时荧光定量PCR反应体系,并用该体系对呼吸道相关致病菌进行检测.鉴定结果与临床常规鉴定方法对照,评价其敏感度、特异度及重复性.结果 通过对100例样品的检测,结果显示该双重实时荧光定量PCR法检测标本的鉴定结果与常规鉴定方法的结果对照,特异度为100%,敏感度为100%;最小能检测到10个拷贝数的重组质粒;批内重复实验和批间重复实验结果均与常规鉴定方法结果相符.结论 双重荧光定量PCR法鉴定克柔念珠菌和光滑念珠菌,特异度和敏感度高,重复性好,且快速、简便,该方法将有助于念珠菌病的早期诊断和针对性治疗.
目的 建立一種快速、靈敏、特異的鑒定剋柔唸珠菌和光滑唸珠菌的雙重實時熒光定量PCR方法.方法 以覈糖體基因內轉錄間隔區Ⅱ(ITSⅡ)為靶目標,設計併閤成分彆針對剋柔唸珠菌、光滑唸珠菌的種特異引物和探針.建立雙重實時熒光定量PCR反應體繫,併用該體繫對呼吸道相關緻病菌進行檢測.鑒定結果與臨床常規鑒定方法對照,評價其敏感度、特異度及重複性.結果 通過對100例樣品的檢測,結果顯示該雙重實時熒光定量PCR法檢測標本的鑒定結果與常規鑒定方法的結果對照,特異度為100%,敏感度為100%;最小能檢測到10箇拷貝數的重組質粒;批內重複實驗和批間重複實驗結果均與常規鑒定方法結果相符.結論 雙重熒光定量PCR法鑒定剋柔唸珠菌和光滑唸珠菌,特異度和敏感度高,重複性好,且快速、簡便,該方法將有助于唸珠菌病的早期診斷和針對性治療.
목적 건립일충쾌속、령민、특이적감정극유념주균화광활념주균적쌍중실시형광정량PCR방법.방법 이핵당체기인내전록간격구Ⅱ(ITSⅡ)위파목표,설계병합성분별침대극유념주균、광활념주균적충특이인물화탐침.건립쌍중실시형광정량PCR반응체계,병용해체계대호흡도상관치병균진행검측.감정결과여림상상규감정방법대조,평개기민감도、특이도급중복성.결과 통과대100례양품적검측,결과현시해쌍중실시형광정량PCR법검측표본적감정결과여상규감정방법적결과대조,특이도위100%,민감도위100%;최소능검측도10개고패수적중조질립;비내중복실험화비간중복실험결과균여상규감정방법결과상부.결론 쌍중형광정량PCR법감정극유념주균화광활념주균,특이도화민감도고,중복성호,차쾌속、간편,해방법장유조우념주균병적조기진단화침대성치료.
Objective To set up a duplex quantitative real-time PCR (QPCR) assay with high sensitivity, specificity and rapidity to detect Candida krusei and Candida glabrata. Methods A pair of species-specific primers, combined with two probes which were selected from the internally transcribed spacers Ⅱ (ITS Ⅱ), were used to amplify the common pathogen associated with the respiratory system in order to evaluate its sensitivity, specificity and repeatability. Results The results obtained from the duplex QPCR assay and the general culture were the same by detecting the samples of 100 cases. Its specificity was 100%,and its sensitivity was 100%. The lower limit of detection of this duplex QPCR assay was 10 copys of recombinant plasmid DNA. The result within group and between groups were the same as the general culture.Conclusion The duplex quantitative real-time PCR assay can detect Candida krusei and Candida glabrata sensitively, specifically, rapidly, simply and stably, and is useful to early diagnosis and target treatment of the diseases caused by the Candida.
