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
2009年
1期
79-83
,共5页
田国忠%张砺%任红宇%王晓蕾%李晋蜀%李晓静%高源%李学春%郑玉红%邓克芬%崔志刚%邵祝军
田國忠%張礪%任紅宇%王曉蕾%李晉蜀%李曉靜%高源%李學春%鄭玉紅%鄧剋芬%崔誌剛%邵祝軍
전국충%장려%임홍우%왕효뢰%리진촉%리효정%고원%리학춘%정옥홍%산극분%최지강%소축군
流感嗜血杆菌%生物学分型%血清学分型%脉冲场凝胶电泳
流感嗜血桿菌%生物學分型%血清學分型%脈遲場凝膠電泳
류감기혈간균%생물학분형%혈청학분형%맥충장응효전영
Haemophilus influenzae%Biotyping%Scrotyping%Pulsed-field gel electrophoresis
目的 应用生物学分型、血清学分型和脉冲场凝胶电泳技术(PFGE)分析临床分离流感嗜血杆菌的表型和分子分型特征,了解流感嗜血杆菌的分子流行病学规律.方法 1988年和2004-2007年期间,成都市儿童医院从患者的下呼吸道分泌物分离培养273株流感嗜血杆菌,菌株的分离培养与鉴定参照美国<临床微生物手册>.菌株的生物学分型和血清学分型分别参照Kilian和Pittman分类法.应用PCR方法对所有的菌株进行荚膜鉴定和血清学分型鉴定.随机选择100株流感嗜血杆菌菌株,应用PFGE技术进行分子分型研究.结果 273株流感嗜血杆菌可分为8个生物型.17.6%(48/273)的菌株为生物型I型,43.6%(119/273)为生物型Ⅱ型,22.7%(62/273)为生物型Ⅲ型,7.3%(20/273)为生物型Ⅳ型,5.9%(16/273)为生物型V型,0.4%(1/273)为生物型Ⅵ型,1.8%(5/273)为生物型Ⅶ型,0.7%(2/273)为生物型Ⅷ型.99.6%(272/273)的菌株为无荚膜的、血清学不可分型的流感嗜血杆菌,1株为血清型f型.100株流感嗜血杆菌经PFGE分析,可分为96个PFGE基因型,93株菌株各自代表一个PFGE基因型.基因型的分布与菌株的分离时间无明显的相关性.结论 引起儿童下呼吸道感染的流感嗜血杆菌主要是无荚膜的、不可分型流感嗜血杆菌菌株,生物型主要是Ⅰ、Ⅱ和Ⅲ型.血清学不可分型流感嗜血杆菌菌株PFGE基因型呈现明显的多态性,菌株的分子多态性可能是流感嗜血杆菌感染在成都市没有形成暴发与流行的分子流行病学原因;PFGE基因分型与生物学分型和血清学分型比较,具有很强的菌株分辨能力,是流感嗜血杆菌感染流行病学研究适用的分析方法.
目的 應用生物學分型、血清學分型和脈遲場凝膠電泳技術(PFGE)分析臨床分離流感嗜血桿菌的錶型和分子分型特徵,瞭解流感嗜血桿菌的分子流行病學規律.方法 1988年和2004-2007年期間,成都市兒童醫院從患者的下呼吸道分泌物分離培養273株流感嗜血桿菌,菌株的分離培養與鑒定參照美國<臨床微生物手冊>.菌株的生物學分型和血清學分型分彆參照Kilian和Pittman分類法.應用PCR方法對所有的菌株進行莢膜鑒定和血清學分型鑒定.隨機選擇100株流感嗜血桿菌菌株,應用PFGE技術進行分子分型研究.結果 273株流感嗜血桿菌可分為8箇生物型.17.6%(48/273)的菌株為生物型I型,43.6%(119/273)為生物型Ⅱ型,22.7%(62/273)為生物型Ⅲ型,7.3%(20/273)為生物型Ⅳ型,5.9%(16/273)為生物型V型,0.4%(1/273)為生物型Ⅵ型,1.8%(5/273)為生物型Ⅶ型,0.7%(2/273)為生物型Ⅷ型.99.6%(272/273)的菌株為無莢膜的、血清學不可分型的流感嗜血桿菌,1株為血清型f型.100株流感嗜血桿菌經PFGE分析,可分為96箇PFGE基因型,93株菌株各自代錶一箇PFGE基因型.基因型的分佈與菌株的分離時間無明顯的相關性.結論 引起兒童下呼吸道感染的流感嗜血桿菌主要是無莢膜的、不可分型流感嗜血桿菌菌株,生物型主要是Ⅰ、Ⅱ和Ⅲ型.血清學不可分型流感嗜血桿菌菌株PFGE基因型呈現明顯的多態性,菌株的分子多態性可能是流感嗜血桿菌感染在成都市沒有形成暴髮與流行的分子流行病學原因;PFGE基因分型與生物學分型和血清學分型比較,具有很彊的菌株分辨能力,是流感嗜血桿菌感染流行病學研究適用的分析方法.
목적 응용생물학분형、혈청학분형화맥충장응효전영기술(PFGE)분석림상분리류감기혈간균적표형화분자분형특정,료해류감기혈간균적분자류행병학규률.방법 1988년화2004-2007년기간,성도시인동의원종환자적하호흡도분비물분리배양273주류감기혈간균,균주적분리배양여감정삼조미국<림상미생물수책>.균주적생물학분형화혈청학분형분별삼조Kilian화Pittman분류법.응용PCR방법대소유적균주진행협막감정화혈청학분형감정.수궤선택100주류감기혈간균균주,응용PFGE기술진행분자분형연구.결과 273주류감기혈간균가분위8개생물형.17.6%(48/273)적균주위생물형I형,43.6%(119/273)위생물형Ⅱ형,22.7%(62/273)위생물형Ⅲ형,7.3%(20/273)위생물형Ⅳ형,5.9%(16/273)위생물형V형,0.4%(1/273)위생물형Ⅵ형,1.8%(5/273)위생물형Ⅶ형,0.7%(2/273)위생물형Ⅷ형.99.6%(272/273)적균주위무협막적、혈청학불가분형적류감기혈간균,1주위혈청형f형.100주류감기혈간균경PFGE분석,가분위96개PFGE기인형,93주균주각자대표일개PFGE기인형.기인형적분포여균주적분리시간무명현적상관성.결론 인기인동하호흡도감염적류감기혈간균주요시무협막적、불가분형류감기혈간균균주,생물형주요시Ⅰ、Ⅱ화Ⅲ형.혈청학불가분형류감기혈간균균주PFGE기인형정현명현적다태성,균주적분자다태성가능시류감기혈간균감염재성도시몰유형성폭발여류행적분자류행병학원인;PFGE기인분형여생물학분형화혈청학분형비교,구유흔강적균주분변능력,시류감기혈간균감염류행병학연구괄용적분석방법.
objective To investigate the epidemiological and molecular typing features of the pathogenic Haemophilus influenzae(H.influenzae)by biotyping,serotyping and pulsed-field gel electrophoresis(PFGE).Methods A total of 273 invasive isolates of H influenzae were collected from the pediatric patients with pneumonia at Chengdu Children Hospital of Sichuan province from 1988 and 2004 to 2007.The idenbfication of H.influenzae strains were done according to the laboratory standard methodology described by Manual of Clinical Microbiology(American).All strains were biotyped according to Kilian's classification with the API[R]NH system.And serotyped by a slide agglutination assay with type a to f specific antlaerum as described by Pittman.PCR method for identification of H.influenzae were performed as described by Falla.One hundred of 273 strains were analyzed by PFGE as described by Saito with some modifications.The resuIts of PFGE were analyzed by Bionumerics soft(Version 4.0,Applied Maths BVBA,Belium).Restilts 78.2%of 273 cases occurred under 1 years old.Eight biotypes were found among the 273 H.influenzae isolates.17.6%(48/273)of all isolates belonged to biotype Ⅰ,43.6%(119/273)were biotype Ⅱ,22.7%(62/273)were biotype Ⅲ,7.3%(20/273)were biotype Ⅳ,5.9%(16/273)were biotype Ⅴ,0.4%(1/273)were biotype Ⅵ,1.8%(5/273)were biotype Ⅶ and 0.7%(2/273)were biotype Ⅷ.respeetively.99.6% of all 273 isolates were nontypeable.There was only one isolate was serotvpe f Ninty-six PFGE genotypes were obtained in this study.One hundred strains demonstrated a variety of genomic Datterns by PFGE.The most isolates of the flame PFGE genotype(type 35)was 3 isolates.Each of93 PFGE genotypes was represented by only a single isolate.The genotypes distribution didn't correlate with the time distribution of the strains were isolated.Conclusion Nontypeable H.influenzae primarily caused acute Dneumoma in children under 1 years old.They mostly belonged to biotype Ⅰ,Ⅱ and Ⅲ biotypes.The nontypeable H.influenzae strains appeared to more heterogeneous patterns by PFGE genotyping.Genotyping may helP understand the molecular characteristics of outbreak and endemicity according to the results of PFGE.PFGE genotyping proved to have a much stronger discriminatory power than either serotyping or biotyping.Our findings suggest that PFGE analysis is useful for the epidemiologieal study of H.influenzae infections.
