中华皮肤科杂志
中華皮膚科雜誌
중화피부과잡지
Chinese Journal of Dermatology
2011年
11期
779-782
,共4页
彭锐锐%尹跃平%魏万惠%王红春%张津萍%陈祥生
彭銳銳%尹躍平%魏萬惠%王紅春%張津萍%陳祥生
팽예예%윤약평%위만혜%왕홍춘%장진평%진상생
梅毒%密螺旋体%苍白%基因分型
梅毒%密螺鏇體%蒼白%基因分型
매독%밀라선체%창백%기인분형
Syphilis%Treponema pallidum%Molecular typing
目的 分析运用最新的三基因定位分型方法检测梅毒螺旋体基因型别的敏感性与特异性.方法 分别采用Nichols标准株和快速血浆反应素环状卡片试验(RPR)、梅毒螺旋体明胶凝集试验(TPPA)均阴性的生殖器疱疹患者湿润性溃疡皮损DNA提取液作为阳性和阴性标本.分析arp基因60个碱基对重复序列的数目、tprEGJ基因MseI酶切后限制性片段长度多态性的型别和tp0548基因序列的型别,根据上述三基因的分析结果,分析梅毒螺旋体基因型别.临床标本来自一期或二期梅毒患者的湿润性皮损.先经梅毒螺旋体特异的polA基因扩增,阳性者用建立的分型方法进行分析.结果 Nichols标准株的基因型别为14a/a,阴性对照的三基因均无扩增.临床标本中三个基因的扩增敏感性分别是94.1％、91.2％和94.1％；91.2％的临床标本检测出完整的基因型别；3个基因的优势型别分别是14型、d型和f型.结论 改良的三基因分型方法检测梅毒螺旋体基因型别具有敏感性高、特异性好、区分度强的特点.
目的 分析運用最新的三基因定位分型方法檢測梅毒螺鏇體基因型彆的敏感性與特異性.方法 分彆採用Nichols標準株和快速血漿反應素環狀卡片試驗(RPR)、梅毒螺鏇體明膠凝集試驗(TPPA)均陰性的生殖器皰疹患者濕潤性潰瘍皮損DNA提取液作為暘性和陰性標本.分析arp基因60箇堿基對重複序列的數目、tprEGJ基因MseI酶切後限製性片段長度多態性的型彆和tp0548基因序列的型彆,根據上述三基因的分析結果,分析梅毒螺鏇體基因型彆.臨床標本來自一期或二期梅毒患者的濕潤性皮損.先經梅毒螺鏇體特異的polA基因擴增,暘性者用建立的分型方法進行分析.結果 Nichols標準株的基因型彆為14a/a,陰性對照的三基因均無擴增.臨床標本中三箇基因的擴增敏感性分彆是94.1％、91.2％和94.1％；91.2％的臨床標本檢測齣完整的基因型彆；3箇基因的優勢型彆分彆是14型、d型和f型.結論 改良的三基因分型方法檢測梅毒螺鏇體基因型彆具有敏感性高、特異性好、區分度彊的特點.
목적 분석운용최신적삼기인정위분형방법검측매독라선체기인형별적민감성여특이성.방법 분별채용Nichols표준주화쾌속혈장반응소배상잡편시험(RPR)、매독라선체명효응집시험(TPPA)균음성적생식기포진환자습윤성궤양피손DNA제취액작위양성화음성표본.분석arp기인60개감기대중복서렬적수목、tprEGJ기인MseI매절후한제성편단장도다태성적형별화tp0548기인서렬적형별,근거상술삼기인적분석결과,분석매독라선체기인형별.림상표본래자일기혹이기매독환자적습윤성피손.선경매독라선체특이적polA기인확증,양성자용건립적분형방법진행분석.결과 Nichols표준주적기인형별위14a/a,음성대조적삼기인균무확증.림상표본중삼개기인적확증민감성분별시94.1％、91.2％화94.1％；91.2％적림상표본검측출완정적기인형별；3개기인적우세형별분별시14형、d형화f형.결론 개량적삼기인분형방법검측매독라선체기인형별구유민감성고、특이성호、구분도강적특점.
Objective To evaluate the performance of a three-gene typing system in the determination of Treponema pallidum (Tp) genotypes.Methods To determine the genotypes of Tp,three targets were assessed,including the number of 60 base-pair repeats,restriction fragment length polymorphism (RFLP) pattem of tprEGJ gene after MseI digestion and the sequence of tp0548 gene.The DNA extracted from the Nichols strain of Tp served as the positive control,and that from the moist ulcer of patients with genital herpes and negative RPR or TPPA test results served as the negative control.To validate the typing method,clinical specimens were collected from the moist skin lesions of patients with primary or secondary syphilis,and subjected to the amplification of polA gene by PCR.The enhanced molecular typing system was used to determine the genotypes of Tp in Tp DNA-positive specimens.Results The Nichols strain harbored a genotype of 14a/a.No amplification of any of the three target genes was found in the negative control.The arp gene,tprEGJ gene and tp0548 gene were amplified from 94.1％,91.2％ and 94.1％ of the 40 clinical specimens,and the genotype was successfully determined by the three-gene typing system for 91.2％ of the clinical Tp strains.The predominant type of arp,tprEGJ and tp0548 genes was 14 repeats,d and f,respectively in these clinical Tp isolates.Conclusion The enhanced molecular tying method for Tp exhibits high sensitivity,specificity and discrimination potential.
