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

【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~ Ⅳ 4个亚型，TPA分为Ⅴ~ Ⅹ 6个亚型，TPA临床株分为Ⅶ、Ⅸ、Ⅹ、Ⅺ 4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。

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

【Abstract】 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 of Ⅰ- Ⅳ, TPA strains were divided into 6 subtypes of Ⅴ-Ⅹ, and TPA clinical strains were classified into 4 subtypes of Ⅶ, 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.