Sequence of cDNA coding for a 65 kDa adhesive protein for the specific detection of Trichomonas vaginalis by PCR.
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.
Rappelli
P
P
Istituto di Microbiologia e Virologia, Università di Sassari, Italy.
Rocchigiani
A M
AM
Erre
G
G
Colombo
M M
MM
Cappuccinelli
P
P
Fiori
P
P
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D018076:DNA, Complementary / Q000737:chemistry*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D015800:Protozoan Proteins / Q000235:genetics*; D014246:Trichomonas vaginalis / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity
D017931:DNA Primers; D018076:DNA, Complementary; D015800:Protozoan Proteins