Detección de Chlamydia trachomatis en orina por LCR: aplicación del método de mezcla de muestras biológicas / Ligase chain reaction testing of pooled urine specimens to diagnose Chlamydia trachomatis infection

Objective. The purpose of this study was to assess the utility and validity of pooling urine samples for molecular diagnosis of Chlamydia trachomatis infection. Material and methods. Of 1,220 urine samples collected from Mexican female and male adolescents, 305 pools were composed of fourth individual samples each, based on a calculation of optimal pool size. These were processed by ligase chain reaction (LCR) for the detection of C. trachomatis. Positive and gray-zone pools were reanalyzed individually. Cost savings were calculated comparing actual costs of testing to the cost that would have been incurred testing all 1,220 samples individually. Results.Pools results were: 56 positive, 19 gray-zones and 230 negative. Following individual retesting of positive and gray-zone pools, 59 cases of C. trachomatis infection were identified (4.8% prevalence). Thus, a total of 601 LCR tests were performed, for a 50.4% savings considering only the direct cost of the test. Conclusions.Our experience shows that sample pooling is both a reliable and convenient tool for CT surveillance in our setting. It should be considered in other similar settings where limited resources constraint surveillance of STIs.

Objetivo. Evaluar la validez y conveniencia de la estrategia de la mezcla de muestras de orinas para el diagnóstico molecular de Chlamydia trachomatis (CT). Material y métodos. A partir de 1,220 muestras de orina recolectadas de jóvenes de uno y otro sexos, se conformaron 305 mezclas con cuatro alícuotas de muestras individuales, previo cálculo del tamaño óptimo de la mezcla. A continuación se determinó la presencia de ácidos nucleicos de clamidia en esas mezclas, mediante el método de reacción en cadena de la ligasa. Las mezclas positivas o en zona gris fueron reanalizadas de manera individual (cuatro pruebas adicionales). El número final de pruebas realizadas se comparó con el total de pruebas que se habrían efectuado individualmente. Resultados. Del total de mezclas analizadas, 230 resultaron negativas, 56 fueron positivas y 19 más se ubicaron en zona gris. Una vez reanalizadas de manera individual las mezclas positivas y las de zona gris, se obtuvieron 59 muestras de orina positivas a clamidia (prevalencia de 4.81%). De esta manera, el número total de pruebas efectuadas fue de 605 en contraste con las 1,220 que tendrían que haberse hecho si se hubieran procesado las muestras individualmente, es decir, que se logró un ahorro de 50.5% del costo directo del reactivo de diagnóstico. Conclusiones. La metodología aplicada mostró ser tanto confiable como conveniente en el entorno mexicano para llevar a cabo vigilancia epidemiológica de la infección por CT. Dado lo anterior, esta metodología podría ser considerada en otros entornos en los que la falta de recursos limita la vigilancia de las infecciones de transmisión sexual.

Infección por Chlamydia trachomatis en hombres chilenos asintomáticos y con uretritis: utilidad de la orina de primer chorro / Chlamydia trachomatis infection in asymtomatic men with urethritis: usefulness of first catch urine samples

Background: Chlamydia trachomatis is responsible for 30 to 50 percent of genital tract infections and is present, without symptoms, in 20 percent of men and 60 percent of women. We have little information in Chile about the prevalence of ths infection. Aim: to assess the prevalence of Chlamydia trachomatis in men, using first catch urine samples. Subjects and methods: three hundred and fifty one first catch urine samples of asymptomatic men and 50 samples coming from men with a primary urethritis, were analyzed. Urethral discharge samples from the latter were simultaneously studied. Analysis was performed using an enzyme immunoanalysis (MicroTrak Chlamydia EIA, Syva Co.) and a nested polymerase chain reaction towards the gene that codifies MOMP (PCR/OMP). Results: amoung asymptomatic men, two of 154 teenagers aged 18 to 19 years old (1.3 percent), 10 of 100 university students (10 percent) and eight of 97 adults over 30 years old (8.2 percent), were infected. The global prevalence of infection in these men was 5.7 percent. The prevalence of infection i men with urethritis was 12 percent. Urine EIA had a higher detection frequency than PCR/OMP, but according to another PCR assay, these results were false positives. EIA in first catch urine, had a sensitivity, specificity, positive and negative predictive value of 83.3, 75, 31.3 and 97 percent respectively, for the detection of Chlamydia trachomatis. The figures for PCR/OMP were 100 percent for all these parameters. Conclusions: the prevalence of Chlamydia trachomatis infection in Chilean men is similar to that reported in developed countries. Enzyme immuno assay in first catch urine had a good diagnostic accuracy and could be used in epidemiological studies in asymptomatic men