Fluorescence in situ hybridization for identification of Tritrichomonas foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomonosis

被引:9
作者
Gookin, J. L. [1 ]
Stone, M. R. [1 ]
Yaeger, M. J. [2 ]
Meyerholz, D. K. [3 ]
Moisan, Peter [4 ]
机构
[1] N Carolina State Univ, Dept Clin Sci, Coll Vet Med, Raleigh, NC 27695 USA
[2] Iowa State Univ, Coll Vet Med, Ames, IA USA
[3] Univ Iowa, Carver Coll Med, Dept Pathol, Iowa City, IA USA
[4] Rollins Anim Dis Diagnost Lab, Raleigh, NC USA
关键词
Trichomonas; Intestine; Identification; Paraffin; Fluorescence; PENTATRICHOMONAS-HOMINIS; RNA; AUTOFLUORESCENCE; INFECTION; TISSUES; CATS;
D O I
10.1016/j.vetpar.2010.04.014
中图分类号
R38 [医学寄生虫学]; Q [生物科学];
学科分类号
090105 [作物生产系统与生态工程]; 100103 [病原生物学];
摘要
In the present study a highly species-specific oligonucleotide sequence of Tritrichomonas foetus 185 rRNA was used to design an antisense probe for identification of T foetus in formalin-fixed and paraffin-embedded histological specimens by means of fluorescence in Situ hybridization (FISH) Using archival histological specimens from several species with light microscopic evidence of intestinal trichomonosis and under optimized hybridization conditions the probe positively identified trichomonads in colonic specimens from piglets and a kitten with PCR-confirmed T foetus infection Neither positive hybridization of the probe or PCR amplification of T foetus DNA was observed in histological specimens from hamster (Tritrichomonas muris) turkey nor mouse (Entamoeba muris) intestinal protozoal infections Sequence-specific binding of the probe was further verified by successfully out-competing the hybridization with 10 x molar excess unlabeled probe and failure of a labeled sense probe to hybridize The FISH assay described here enables simultaneous location and molecular identification of T foetus in formalin-fixed and paraffin-embedded histological specimens of intestinal trichomonosis The methods employed are likely to also be applicable to probes designed for specific recognition of other trichomonad species especially in mammalian tissue where red blood cell auto-fluorescence can be easily differentiated from the hybridization signal of trichomonads (C) 2010 Elsevier B V All rights reserved
引用
收藏
页码:139 / 143
页数:5
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