Open AccessSequence of cDNA coding for a 65 kDa adhesive protein for the specific detection of Trichomonas vaginalis by PCR
Author(s) -
Rappelli Paola,
Rocchigiani Angela M.,
Erre Giuseppe,
Colombo Mauro M.,
Cappuccinelli Piero,
Fiori PierLuigi
Publication year - 1995
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1995.tb07551.x
Subject(s) - trichomonas vaginalis , biology , amplicon , complementary dna , polymerase chain reaction , microbiology and biotechnology , gene , trichomonas , recombinant dna , coding region , sequence analysis , nucleic acid sequence , cdna library , genetics
A Trichomonas vaginalis cDNA library was constructed and recombinant plaques were screened using rabbit immunoglobulins specific for P65, a protozoan protein involved in pathogenicity that we identified in a previous study. A 1.38 kilobases cDNA fragment coding for the P65 protein was cloned in E. coli and then sequenced. On the basis of of the sequence obtained, six primers were synthesised and used to set up a Polymerase Chain Reaction. The presence of a specific amplicon in all 30 clinical isolates tested shows that P65 is a conserved and stable gene. The reaction is highly sensitive (as few as 5 to 10 parasites can be detected) and specific for Trichomonas vaginalis ; the gene coding P65 adhesin can be therefore considered a very good molecular target for polymerase chain reaction‐based diagnostic purposes.