上海农业学报
上海農業學報
상해농업학보
ACTA AGRICULTURAE SHANGHAI
2015年
4期
34-39
,共6页
胡瑞丽%任方方%吴洋洋%许燕%赵凯%索玉娟%邵毅%周昌艳
鬍瑞麗%任方方%吳洋洋%許燕%趙凱%索玉娟%邵毅%週昌豔
호서려%임방방%오양양%허연%조개%색옥연%소의%주창염
多重 PCr%食源性致病菌%检测%食品安全
多重 PCr%食源性緻病菌%檢測%食品安全
다중 PCr%식원성치병균%검측%식품안전
Multiple PCr%Foodborne pathogenic bacteria%Detection%Food safety
根据单核增生李斯特菌、沙门氏菌、金黄色葡萄球菌、大肠杆菌 O157：H7的特异性序列分别设计4对特异性引物，建立了一种快速检测上述4种常见食源性致病菌的多重 PCr 方法，并对反应体系中的模板、引物、rTaq 酶、MgSO4添加量以及退火温度进行了优化。结果显示：4对引物序列的特异性均较好，利用单重 PCr对4种致病菌的灵敏度进行检测，其检测限均达到103 copies/mL。多重 PCr 同时检测4种致病菌的灵敏度为103 copies/mL，对人工污染猪肉中的4种致病菌的灵敏度检测为103 cfu/mL。该检测方法不与其他菌产生交叉反应，与传统方法相比大大缩短了检测时间，并提高了检测的准确度。
根據單覈增生李斯特菌、沙門氏菌、金黃色葡萄毬菌、大腸桿菌 O157：H7的特異性序列分彆設計4對特異性引物，建立瞭一種快速檢測上述4種常見食源性緻病菌的多重 PCr 方法，併對反應體繫中的模闆、引物、rTaq 酶、MgSO4添加量以及退火溫度進行瞭優化。結果顯示：4對引物序列的特異性均較好，利用單重 PCr對4種緻病菌的靈敏度進行檢測，其檢測限均達到103 copies/mL。多重 PCr 同時檢測4種緻病菌的靈敏度為103 copies/mL，對人工汙染豬肉中的4種緻病菌的靈敏度檢測為103 cfu/mL。該檢測方法不與其他菌產生交扠反應，與傳統方法相比大大縮短瞭檢測時間，併提高瞭檢測的準確度。
근거단핵증생리사특균、사문씨균、금황색포도구균、대장간균 O157：H7적특이성서렬분별설계4대특이성인물，건립료일충쾌속검측상술4충상견식원성치병균적다중 PCr 방법，병대반응체계중적모판、인물、rTaq 매、MgSO4첨가량이급퇴화온도진행료우화。결과현시：4대인물서렬적특이성균교호，이용단중 PCr대4충치병균적령민도진행검측，기검측한균체도103 copies/mL。다중 PCr 동시검측4충치병균적령민도위103 copies/mL，대인공오염저육중적4충치병균적령민도검측위103 cfu/mL。해검측방법불여기타균산생교차반응，여전통방법상비대대축단료검측시간，병제고료검측적준학도。
According to the specific sequence of 4 common foodborne pathogenic bacteria,Listeria monocy-togenes,Salmonella enterica,Staphylococcus aureus and Escherichia coli O157:H7,4 pairs of specific primers were designed and a multiple PCr method for rapid detecting the above bacteria was established.The additive amount of template,primers,rTaq,MgSO4 ,and the annealing temperature of the reaction system were optimized.The re-sults showed that the specificity of 4 pairs of primers was better.The sensitivity of 4 species of pathogenic bacteria was detected by single PCr,and the detection limit was 103 copies/mL.The sensitivity of multiple PCr for simul-taneous detection of 4 species of pathogenic bacteria was 103 copies/mL,and the sensitivity of 4 species of patho-genic bacteria in artificial contaminated pork was detected as 103 cfu/mL.The detection method did not produce cross reactions with other bacteria.Compared with the traditional method,the detection time was greatly short-ened,and the accuracy of detection was improved.
