基于TP0136蛋白异质性建立一种新的苍白密螺旋体分子分型方法  

Establishment of a new molecular typing method for Treponema pallidum based on TP0136 protein sequence heterogeneity

作　　者：魏然 柯吴坚 陈文韬 谭玲俏 刘雅慧 吕萍 黄涛 张君 张晓辉 王柳苑 车雅敏[1] Wei Ran;Ke Wujian;Chen Wentao;Tan Lingqiao;Liu Yahui;Lyu Ping;Huang Tao;Zhang Jun;Zhang Xiaohui;Wang Liuyuan;Che Yamin(Department of Dermatology and Venereology,General Hospital,Tianjin Medical University,Tianjin 300052,China;The Second Department of Dermatology,Dermatology Hospital of Southern Medical University,Guangzhou 510091,China;Department of Dermatology and Venereology,Yangjiang Public Health Hospital,Yangjiang 529500,Guangdong,China;Department of Dermatology and Venereology,Chronic Disease Prevention Hospital of Qingyuan City,Qingyuan 511500,Guangdong,China)

机构地区：[1]天津医科大学总医院皮肤性病科,300052 [2]南方医科大学皮肤病医院皮肤二科,广州510091 [3]阳江市公共卫生医院皮肤性病科,广东529500 [4]清远市慢性病防治医院皮肤性病科,广东511500

出　　处：《中华皮肤科杂志》2020年第7期546-550,共5页Chinese Journal of Dermatology

基　　金：国家自然科学基金(81772240);广东省医学科研基金(A2018248、A2017224);广州市健康医疗协同创新重大专项(201704020219);南方医科大学皮肤病医院基础研究苗圃项目(C2019001);深圳龙华区引进高层次医学团队项目(HLPM201907020105)。

摘　　要：目的基于苍白密螺旋体属(TP)TP0136蛋白的异质性,建立一种新的TP分子分型方法。方法从GenBank中查询9株梅毒苍白密螺旋体(TPA)、3株雅司苍白密螺旋体(TPE)、1株未分类的类人猿密螺旋体(FB)和1株地方密螺旋体(TEN)的TP0136开放阅读框(ORF)氨基酸序列并进行多重序列比对,得出TP0136蛋白分子分型结果。利用建立的TP0136蛋白分子分型方法,对2015年1月至2018年12月南方医科大学皮肤病医院收集的23株TPA临床分离株进行分类,并将分型结果与传统的Arp/Tpr/TP0548三基因分型方法进行比较。结果TP0136蛋白在不同种TP株中具有高度异质性,根据TP0136氨基酸序列,TPE、FB和TEN分为Ⅰ~Ⅳ4个亚型,TPA分为Ⅴ~Ⅹ6个亚型,TPA临床株分为Ⅶ、Ⅸ、Ⅹ、Ⅺ4个亚型。通过TPA传统的三基因分型方法可将23株TPA临床株分为13D/d、14D/f、14D/g、15D/f、16A/e 5型。将TP0136分型法与传统分型法相结合,可将上述临床株进一步细分为10型,其中14D/f型应用TP0136分型法可进一步分为3个亚型。结论TP0136分子分型方法可用于TP种的区分,有助于传统TPA分子分型的进一步完善。Objective To establish a new molecular typing method for Treponema pallidum(TP)based on TP0136 protein sequence heterogeneity.Methods The amino acid sequences of TP0136 open reading frame(ORF)of 9 strains of Treponema pallidum ssp.Pallidum(TPA),3 strains of Treponema pallidum ssp.Pertenue(TPE),1 unclassified simian strain of Treponema Fribourg-Blanc(FB)and 1 strain of Treponema pallidum ssp.Endemicum(TEN)were searched from Genbank,and multiple sequence comparisons were performed to obtain the molecular typing results of TP0136 protein.The TP0136 protein-based molecular typing method was used to classify 23 TPA clinical isolates,which were collected from Dermatology Hospital of Southern Medical University from January 2015 to December 2018,and the typing results were compared with those by the traditional typing method based on the tp0548/Arp/Tpr genes.Results TP0136 protein was highly heterogeneous in different TP strains.According to the amino acid sequence of TP0136,TPE,FB and TEN strains were divided into 4 subtypes ofⅠ-Ⅳ,TPA strains were divided into 6 subtypes ofⅤ-Ⅹ,and TPA clinical strains were classified into 4 subtypes ofⅦ,Ⅸ,Ⅹ,Ⅺ.Through the traditional typing method described above,23 TPA clinical strains could be divided into 5 types(13D/d,14D/f,14D/g,15D/f,16A/e).By using the TP0136 protein-based typing method combined with traditional typing method,the above clinical strains could be further subdivided into 10 types,and the 14D/f type could be further divided into 3 subtypes by using the TP0136 protein-based typing method.Conclusion The TP0136 protein-based molecular typing method can be used to distinguish TP species,which is helpful for further improvement of traditional TPA molecular typing.

关 键 词：密螺旋体属 梅毒 基因分型技术 序列分析 蛋白质 TP0136 

分 类 号：R759.1[医药卫生—皮肤病学与性病学]