Use of in vitro cytoadherence assays in the comparative study of the virulence of isolates of Trichomonas vaginalis.
This study describes the use of bromo-deoxiuridine, a non-radioactive analogue of thymidine, to determine the adhesion of 40 T. vaginalis isolates, as compared with the clinical manifestations found in patients. Parasite-labeling was optimized and label detection by immunoassay was carried out. Bromo-deoxiuridine at 10 microM for 16 h produced the highest sensitivity. Once optimized, the assay was able to detect between 3.12 x 10(3) and 4 x 10(5) parasites, with a detection limit of 9.14 x 10(2), which is lower than with the use of [3H]-thymidine. The variation coefficients were 2.65% for repeatability and 3.8% for reproducibility. A correlation coefficient of 0.97 was found between the pattern curves obtained by both labeling procedures. The level of adhesion to HeLa cells was directly proportional to the severity of the clinical manifestations (P < 0.05).
Rojas
Lázara
L
Departamento de Parasitología, Instituto de Medicina Tropical "Pedro Kourí", Autopista Novia del Mediodía km 61/2, Apartado Postal 601, Marianao 13 Ciudad de La Habana, Cuba. lrojas@ipk.sld.cu
Sariego
Idalia
I
Fraga
Jorge
J
Sarría
Carlos A
CA
Sarracent
Jorge
J
Sánchez
Lizet
L
D000293:Adolescent; D000818:Animals; D001681:Biological Assay; D001973:Bromodeoxyuridine / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007118:Immunoassay; D066298:In Vitro Techniques; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D014246:Trichomonas vaginalis / Q000166:cytology* / Q000378:metabolism / Q000472:pathogenicity*; D014774:Virulence
D001973:Bromodeoxyuridine