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Development of Nested Polymerase Chain Reaction with Novel Specific Primers for Detection of Tritrichomonas muris Infection in Laboratory Mice

Authors :
Hongbo Zhang
Nan Zhang
Jianhua Li
Panpan Zhao
Xin Li
Xiaocen Wang
Xu Zhang
Bao Yuan
Fei Gao
Pengtao Gong
Xichen Zhang
Source :
Animals, Vol 13, Iss 20, p 3177 (2023)
Publication Year :
2023
Publisher :
MDPI AG, 2023.

Abstract

A variety of rodent ceca are parasitized by Tritrichomonas muris (T. muris), a flagellated protozoan. To date, there are no ideal methods for the detection of T. muris infections in laboratory mice; thus, new molecular methodologies for its specific detection need to be developed. In this study, using staining and SEM, it was observed that T. muris has a pear-shaped body and contains three anterior flagella. A nested PCR system with novel specific primers was designed based on the conserved regions of the SSU rRNA gene of T. muris. The nested PCR system for T. muris showed good specificity and high sensitivity for at least 100 T. muris trophozoites/mL and 0.1 ng/μL of fecal genomic DNA, which means that 176 trophozoites per gram of mouse feces could be detected. When using this nested PCR system, the detection rate was 18.96% (58/306), which was higher than the detection rate of 14.05% (43/306) detected via smear microscopy in fecal samples from five mouse strains. The sensitivity and specificity of nested PCR in detecting T. muris was found to be 100%, and it demonstrated a 26% increase in diagnostic sensitivity compared to the smear microscopy method in the present study. In conclusion, the nested PCR developed with novel primers based on the SSU rRNA gene of T. muris has good accuracy, specificity, and sensitivity for the detection of T. muris infections in laboratory mice.

Details

Language :
English
ISSN :
13203177 and 20762615
Volume :
13
Issue :
20
Database :
Directory of Open Access Journals
Journal :
Animals
Publication Type :
Academic Journal
Accession number :
edsdoj.80af509eda41cdad47238c69f8a35d
Document Type :
article
Full Text :
https://doi.org/10.3390/ani13203177