中国动物传染病学报
中國動物傳染病學報
중국동물전염병학보
CHINESE JOURNAL OF VETERINARY PARASITOLOGY
2013年
5期
51-56
,共6页
李雅文%刘远佳%郑国超%谭立娉%胡伟%罗琴%林丽琴%路鹏云%李国清
李雅文%劉遠佳%鄭國超%譚立娉%鬍偉%囉琴%林麗琴%路鵬雲%李國清
리아문%류원가%정국초%담립빙%호위%라금%림려금%로붕운%리국청
蓝氏贾第虫%犬%核糖体基因间隔区%PCR检测
藍氏賈第蟲%犬%覈糖體基因間隔區%PCR檢測
람씨가제충%견%핵당체기인간격구%PCR검측
Giardialamblia%dogs%intergenic spacers%PCR
应用PCR技术对广州市某宠物寄养所采集的一株D型犬源贾第虫和一株F型猫源贾第虫的核糖体IGS序列进行了扩增、克隆、测序,将测序结果与GenBank已上传的贾第虫相应序列进行比对分析,基于贾第虫IGS序列建立了具有良好特异性和敏感性的PCR检测方法,并对84份临床粪样进行了检测。结果显示,D型犬源贾第虫和F型猫源贾第虫的IGS序列长分别为1355 bp和1388 bp,贾第虫IGS序列存在多态性现象,种间差异明显,可以作为区分不同基因型的分子标记;建立的PCR方法能特异性扩增犬源贾第虫核糖体IGS序列,而犬蛔虫等对照虫体DNA均不能扩增,该方法对贾第虫DNA的最小检测量为82 fg,对84份临床粪样的检出率为3.57%,比传统镜检法高出2.38%,具有一定的临床应用价值。
應用PCR技術對廣州市某寵物寄養所採集的一株D型犬源賈第蟲和一株F型貓源賈第蟲的覈糖體IGS序列進行瞭擴增、剋隆、測序,將測序結果與GenBank已上傳的賈第蟲相應序列進行比對分析,基于賈第蟲IGS序列建立瞭具有良好特異性和敏感性的PCR檢測方法,併對84份臨床糞樣進行瞭檢測。結果顯示,D型犬源賈第蟲和F型貓源賈第蟲的IGS序列長分彆為1355 bp和1388 bp,賈第蟲IGS序列存在多態性現象,種間差異明顯,可以作為區分不同基因型的分子標記;建立的PCR方法能特異性擴增犬源賈第蟲覈糖體IGS序列,而犬蛔蟲等對照蟲體DNA均不能擴增,該方法對賈第蟲DNA的最小檢測量為82 fg,對84份臨床糞樣的檢齣率為3.57%,比傳統鏡檢法高齣2.38%,具有一定的臨床應用價值。
응용PCR기술대엄주시모총물기양소채집적일주D형견원가제충화일주F형묘원가제충적핵당체IGS서렬진행료확증、극륭、측서,장측서결과여GenBank이상전적가제충상응서렬진행비대분석,기우가제충IGS서렬건립료구유량호특이성화민감성적PCR검측방법,병대84빈림상분양진행료검측。결과현시,D형견원가제충화F형묘원가제충적IGS서렬장분별위1355 bp화1388 bp,가제충IGS서렬존재다태성현상,충간차이명현,가이작위구분불동기인형적분자표기;건립적PCR방법능특이성확증견원가제충핵당체IGS서렬,이견회충등대조충체DNA균불능확증,해방법대가제충DNA적최소검측량위82 fg,대84빈림상분양적검출솔위3.57%,비전통경검법고출2.38%,구유일정적림상응용개치。
The intergenic spacers (IGS) were amplified in PCR from nuclear ribosomal DNAs of two Giardialamblia strains (assemble D from dog and assemble F from cat) isolated from feces of naturally infected pets in Guangzhou. The PCR products were purified, cloned and sequenced. The resulting nucleotide sequences were compared with related sequences in the GenBank databases. The IGS sequences were 1355 bp for the canine Giardia strain and 1388 bp for the feline Giardia strain. The interspecific difference in the IGS sequence might serve as a genetic marker for the identification and differentiation of Giardia lamblia. Furthermore, a PCR detection method was developed based on IGS sequence. The established PCR assay specifically amplified IGS sequence of G. lamblia but not all control parasites such as Toxocaracanis etc. The minimum detection limit of G.lamblia was 82 fg. Total 84 clinical samples were tested using this method. The positive rate of clinical samples was 3.57%, higher than 2.38%by traditional method.