Construction of a cDNA library for Sparganum mansoni and screening of diagnostic antigen cadidates
10.16250/j.32.1374.2021143
- VernacularTitle:曼氏裂头蚴cDNA文库的构建及诊断候选抗原筛选
- Author:
Yan LU
1
;
Jia-Xu CHEN
1
;
Peng SONG
1
;
Hao LI
1
;
Lin AI
1
;
Yu-Chun CAI
1
;
Yan-Hong CHU
1
;
Shao-Hong CHEN
1
Author Information
1. National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention (Chinese Center for Tropical Diseases Research), NHC Key Laboratory of Parasite and Vector Biology, WHO Collaborating Centre for Tropical Diseases, National Center for International Research on Tropical Diseases, Shanghai 200025, China
- Publication Type:Journal Article
- Keywords:
Sparganum mansoni;
cDNA library;
Immunoscreening;
Diagnostic antigen;
SMART technique
- From:
Chinese Journal of Schistosomiasis Control
2021;33(4):380-386
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a cDNA library of Sparganum mansoni and immunoscreen antigen candidates for immunodiagnosis of sparganosis mansoni. Methods Total RNA was extracted from S. mansoni, and reversely transcribed into cDNA, which was ligated into the phage vector. These recombinant vectors were packaged in vitro to construct the SMART cDNA library of S. mansoni. Then, the cDNA library was immunoscreened with sera from patients with sparganosis mansoni to yield positive clones. The inserted fragments of positive clones were sequenced and subjected to homology analyses, and the structure and functions of the coding proteins were predicted. Results The SMATR cDNA library of S. mansoni was successfully constructed. The titer of the cDNA library was 6.25 × 106 pfu/mL, with a recombinant efficiency of 100%, and the mean length of the inserted fragments in the library was larger than 1 100 bp. A total of 12 positive clones were obtained by immunoscreening, and were categorized into Sm-I (Sm60-1), Sm-II (Sm58-1), Sm-III (Sm20-1) and Sm-IV (Sm22-3), with 1 134, 1 063, 883 bp and 969 bp long inserted fragments. Their coding proteins were highly homologous with the Spirometra erinaceieuropaei antigenic polypeptide, cytoplasmic antigen, ribosomal protein S4-like protein and unnamed protein product, respectively. Conclusions A SMART cDNA library of S. mansoni has been successfully constructed and 4 categories of positive clones have been identified, which provides a basis for further studies on diagnostic antigens for sparganosis mansoni.
