中华微生物学和免疫学杂志
中華微生物學和免疫學雜誌
중화미생물학화면역학잡지
CHINESE JOURNAL OF MICROBIOLOGY AND IMMUNOLOGY
2011年
10期
916-921
,共6页
何冬梅%王洪敏%柯昌文%邓小玲%杨杏芬%赖蔚苳%柯碧霞%李柏生%谭海玲
何鼕梅%王洪敏%柯昌文%鄧小玲%楊杏芬%賴蔚苳%柯碧霞%李柏生%譚海玲
하동매%왕홍민%가창문%산소령%양행분%뢰위동%가벽하%리백생%담해령
单增李斯特菌%基因芯片%寡核苷酸探针
單增李斯特菌%基因芯片%寡覈苷痠探針
단증리사특균%기인심편%과핵감산탐침
Listeria monocytogenes%DNA microarray%Oligo probe
目的 研制快速、特异、灵敏的检测单增李斯特菌的基因芯片.方法 选择gyrB、ISR、16S rRNA、23S rRNA、hlyA、iap和prfA作为单增李斯特菌的检测靶基因,研制一种Oligo探针基因芯片,对18个不同种属来源的已知参考生物样品进行检测和鉴定,并且采用对比试验、重复性试验、灵敏度试验和特异性试验对该芯片进行验证评估.结果 通过对比发现IDT合成的70 mer Oligo芯片探针在芯片打印与芯片检测两个方面较优.比较10、40和80 μmol/L 3个Oligo探针点样浓度,结果 显示10 μmol/L的探针点样浓度已能获得很好的芯片检测结果.单增李斯特菌检测芯片具有较好的重复性；样品检测绝对量下限为0.9 ng DNA左右.结论 Oligo基因芯片可以快速准确地检测单增李斯特菌.
目的 研製快速、特異、靈敏的檢測單增李斯特菌的基因芯片.方法 選擇gyrB、ISR、16S rRNA、23S rRNA、hlyA、iap和prfA作為單增李斯特菌的檢測靶基因,研製一種Oligo探針基因芯片,對18箇不同種屬來源的已知參攷生物樣品進行檢測和鑒定,併且採用對比試驗、重複性試驗、靈敏度試驗和特異性試驗對該芯片進行驗證評估.結果 通過對比髮現IDT閤成的70 mer Oligo芯片探針在芯片打印與芯片檢測兩箇方麵較優.比較10、40和80 μmol/L 3箇Oligo探針點樣濃度,結果 顯示10 μmol/L的探針點樣濃度已能穫得很好的芯片檢測結果.單增李斯特菌檢測芯片具有較好的重複性；樣品檢測絕對量下限為0.9 ng DNA左右.結論 Oligo基因芯片可以快速準確地檢測單增李斯特菌.
목적 연제쾌속、특이、령민적검측단증리사특균적기인심편.방법 선택gyrB、ISR、16S rRNA、23S rRNA、hlyA、iap화prfA작위단증리사특균적검측파기인,연제일충Oligo탐침기인심편,대18개불동충속래원적이지삼고생물양품진행검측화감정,병차채용대비시험、중복성시험、령민도시험화특이성시험대해심편진행험증평고.결과 통과대비발현IDT합성적70 mer Oligo심편탐침재심편타인여심편검측량개방면교우.비교10、40화80 μmol/L 3개Oligo탐침점양농도,결과 현시10 μmol/L적탐침점양농도이능획득흔호적심편검측결과.단증리사특균검측심편구유교호적중복성；양품검측절대량하한위0.9 ng DNA좌우.결론 Oligo기인심편가이쾌속준학지검측단증리사특균.
Objective To develop a rapid and sensitive DNA microarray for Listeria monocytogenes detection.Methods A DNA microarray was developed using gyrB,ISR,16S rRNA,23S rRNA,hlyA,iap and prfA as the target genes and tested against 18 different species of known reference for repeatability,sensitivity,and specificity to verify the effectiveness of the chip.Results After testing of samples by the LM array,results show that the 70 mer Oligos synthesized by IDT are superior to the Oligos synthesized by Sagon with respect to both probe spotting or samples detection.The comparison of 3 spotting probe concentrations of 10 μmol/L,40 μmol/L and 80 μmol/L demonstrated that the 10 pmol/L probes result in good detection signals equivalent to the 40 μmol/L and 80 μmol/L probes.The repeatability and sensitivity evaluated by sample testing on the LM array revealed that the chips developed in this study have good repeatability and the lower limit of sample detection is 0.9 ng DNA.The LM array can distinguish clearly and definitively between Listeria and non-Listeria bacteria in the sample.Conclusion The microarray is able to rapidly detect and identify Listeria monocytogenes.
