中华实验眼科杂志
中華實驗眼科雜誌
중화실험안과잡지
CHINESE JOURNAL OF EXPERIMENTAL OPHTHALMOLOGY
2013年
9期
867-869
,共3页
安娜%刘先宁%兰雅娴%陶沙
安娜%劉先寧%蘭雅嫻%陶沙
안나%류선저%란아한%도사
泪囊炎%多聚酶链反应%16S rRNA%基因测序%细菌
淚囊炎%多聚酶鏈反應%16S rRNA%基因測序%細菌
루낭염%다취매련반응%16S rRNA%기인측서%세균
Dacryocystitis%Polymerase chain reaction%16S rRNA%Gene sequence%Bacteria
背景 泪囊炎是眼科常见的感染性疾病,对泪囊炎致病菌检测的金标准为培养法,在培养法的基础上结合分子生物学的检测方法可以更精确地鉴定出致病菌的种属,但目前眼科领域类似的研究资料比较缺乏.目的 采用PCR法扩增细菌核糖体16S rRNA基因序列鉴定泪囊炎病原细菌的种属.方法 取10例质控标准菌样本,利用PCR的加热过程使细菌核酸释放,扩增出细菌16S rRNA基因序列,测序后与GenBank中的基因序列进行比对,鉴定出细菌的种属信息,与生化鉴定法结果进行对比,确定16S rRNA基因检测方法的可靠性.取30例泪囊炎患者泪囊分泌物培养出的细菌,利用上述方法进行病原菌鉴定.结果 10例质控标准菌样本经PCR扩增后其产物序列的鉴定结果与生化鉴定法结果完全一致.30例泪囊炎病原细菌标本中,16S rRNA基因序列法鉴定结果为表皮葡萄球菌13例,沃氏葡萄球菌2例,人葡萄球菌1例,麦氏棒杆菌5例,肺炎链球菌3例,蜡状芽孢杆菌2例,藤黄微球菌1例；卡他莫拉菌1例,奥斯陆莫拉菌1例,铜绿假单胞菌1例.结论 通过PCR扩增细菌核糖体16S rRNA基因序列鉴定泪囊炎病原细菌种属的方法准确性高,特异性强.
揹景 淚囊炎是眼科常見的感染性疾病,對淚囊炎緻病菌檢測的金標準為培養法,在培養法的基礎上結閤分子生物學的檢測方法可以更精確地鑒定齣緻病菌的種屬,但目前眼科領域類似的研究資料比較缺乏.目的 採用PCR法擴增細菌覈糖體16S rRNA基因序列鑒定淚囊炎病原細菌的種屬.方法 取10例質控標準菌樣本,利用PCR的加熱過程使細菌覈痠釋放,擴增齣細菌16S rRNA基因序列,測序後與GenBank中的基因序列進行比對,鑒定齣細菌的種屬信息,與生化鑒定法結果進行對比,確定16S rRNA基因檢測方法的可靠性.取30例淚囊炎患者淚囊分泌物培養齣的細菌,利用上述方法進行病原菌鑒定.結果 10例質控標準菌樣本經PCR擴增後其產物序列的鑒定結果與生化鑒定法結果完全一緻.30例淚囊炎病原細菌標本中,16S rRNA基因序列法鑒定結果為錶皮葡萄毬菌13例,沃氏葡萄毬菌2例,人葡萄毬菌1例,麥氏棒桿菌5例,肺炎鏈毬菌3例,蠟狀芽孢桿菌2例,籐黃微毬菌1例；卡他莫拉菌1例,奧斯陸莫拉菌1例,銅綠假單胞菌1例.結論 通過PCR擴增細菌覈糖體16S rRNA基因序列鑒定淚囊炎病原細菌種屬的方法準確性高,特異性彊.
배경 루낭염시안과상견적감염성질병,대루낭염치병균검측적금표준위배양법,재배양법적기출상결합분자생물학적검측방법가이경정학지감정출치병균적충속,단목전안과영역유사적연구자료비교결핍.목적 채용PCR법확증세균핵당체16S rRNA기인서렬감정루낭염병원세균적충속.방법 취10례질공표준균양본,이용PCR적가열과정사세균핵산석방,확증출세균16S rRNA기인서렬,측서후여GenBank중적기인서렬진행비대,감정출세균적충속신식,여생화감정법결과진행대비,학정16S rRNA기인검측방법적가고성.취30례루낭염환자루낭분비물배양출적세균,이용상술방법진행병원균감정.결과 10례질공표준균양본경PCR확증후기산물서렬적감정결과여생화감정법결과완전일치.30례루낭염병원세균표본중,16S rRNA기인서렬법감정결과위표피포도구균13례,옥씨포도구균2례,인포도구균1례,맥씨봉간균5례,폐염련구균3례,사상아포간균2례,등황미구균1례；잡타막랍균1례,오사륙막랍균1례,동록가단포균1례.결론 통과PCR확증세균핵당체16S rRNA기인서렬감정루낭염병원세균충속적방법준학성고,특이성강.
Background Dacryocystitis is one of the most common infectious eye diseases.The gold standard for the identification of bacteria causing dacryocystitis is bacterial culture.The combination of regular culture method with molecular biology techeniques will generate more reliable results.However,very few research data are available in ophthalmological studies in this area.Objective This study was to identify the genera and species of the dacryocystitis-causing bacteria by PCR amplification of the 16S rRNA sequences.Methods Ten cases of qualified standardized bacteria samples were taken,and the nucleic acids were released in the heating process of the PCR procedure.The 16S rRNA genes were amplified and sequenced,and the genera and species were identified using BLAST from GenBank,and the results were used to compare with the results from biochemical identification to test the reliability of this method.The cultured bacterial species from the lacrimal sac secretions from 30 cases of dacryocystitis patients were identified with the above method.Results The outcome of the PCR identification for the 10 cases of quality control standard bacterial specimens was consistent with the results from the biochemical identification.The identification of the 30 cases of dacryocystitis through sequencing the 16S rRNA revealed there were 13 cases of Staphylococcus epidermidis infection,2 cases of Staphylococcus warneri infection,1 case of Staphylococcus hominis infection,5 cases of Corynebacterium macginleyi infection,3 cases of Streptococcus pneumonia infection,2 cases of Bacillus cereus infection,1 case of Micrococcus luteus infection,1 case of Moraxella catarrhalis infection,1 case of Moraxella osloensis infection and 1 case of Pseudomonas aeruginosa infection.Conclusions Sequencing the 16S rRNA is an accurate and specific way for the identification of the genera and species of bacteria that cause dacryocystitis in patients.This sequencing method is feasible in monitoring a variety of dacryocystitis-causing pathogens.More information and epidemiological statistics about dacryocystitis can be obtained from 16S rRNA sequencing.
