Genotyping of Chlamydia trachomatis from endocervical specimens of infertile Mexican women.

INTRODUCTION: It has been reported in several countries that Chlamydia trachomatis genotypes D, E, and F are the ones more frequently associated with urogenital infections. In Mexico, the prevalence of serovars and genotypes is unknown. MATERIAL AND METHODS: One hundred and fifty-two endocervical swabs were collected from infertile women to test for C. trachomatis. The PCR-based RFLP and automated-sequencing methods of ompA gene was used to identify the C. trachomatis genotypes. Sequences of 891 pb obtained were aligned with currently available chlamydial sequences from GenBank to identify the corresponding genotype. RESULTS: Twenty-four women with infertility (15.8%) were positive for C. trachomatis. According to the RFLP and nucleotide sequences results the most prevalent ompA genotype corresponded to serovar F (n=13 [54.2%]), followed by serovars E (n=2 [8.7%]), G (n=2 [8.7%]), K (n=2 [8.7%]) and LGV (n=2 [8.7%]), while serovars D, H and Ia were less prevalent (all n=1 [4.2%]). None of the patients who were positive to genovar L2 had symptoms of lymphogranuloma venereum (LGV). Nucleotide sequences analysis showed a new genovariant of L2, which was different to L2b to L2f. Mutation points were observed in VS1 domain of Omp A. CONCLUSIONS: In this study the most common genotypes were F. Furthermore, the L2 genovariants were demonstrated in infertile women without signs and symptoms of LGV disease. Presence of point mutations in L2 genotype sequences were seen by which there is a need for further research in order to identify new L2 genetic variants that exist in Latin America.

Genotyping of Chlamydia trachomatis from endocervical specimens of infertile Mexican women

Introduction It has been reported in several countries that Chlamydia trachomatis genotypes D, E, and F are the ones more frequently associated with urogenital infections. In Mexico, the prevalence of serovars and genotypes is unknown. Material and Methods One hundred and fifty-two endocervical swabs were collected from infertile women to test for C. trachomatis. The PCR-based RFLP and automated-sequencing methods of ompA gene was used to identify the C. trachomatis genotypes. Sequences of 891 pb obtained were aligned with currently available chlamydial sequences from GenBank to identify the corresponding genotype. Results Twenty-four women with infertility (15.8%) were positive for C. trachomatis. According to the RFLP and nucleotide sequences results the most prevalent ompA genotype corresponded to serovar F (n=13 [54.2%]), followed by serovars E (n=2 [8.7%]), G (n=2 [8.7%]), K (n=2 [8.7%]) and LGV (n=2 [8.7%]), while serovars D, H and Ia were less prevalent (all n=1 [4.2%]). None of the patients who were positive to genovar L2 had symptoms of lymphogranuloma venereum (LGV). Nucleotide sequences analysis showed a new genovariant of L2, which was different to L2b to L2f. Mutation points were observed in VS1 domain of Omp A.Conclusions In this study the most common genotypes were F. Furthermore, the L2 genovariants were demonstrated in infertile women without signs and symptoms of LGV disease. Presence of point mutations in L2 genotype sequences were seen by which there is a need for further research in order to identify new L2 genetic variants that exist in Latin America (AU)

Introducción En diversos países se ha informado que los genotipos de Chlamydia trachomatis más frecuentes y que están asociados a infecciones urogenitales son: D, E y F. En México, la prevalencia de los serotipos y genotipos no se conoce. Material y métodos Se obtuvieron ciento cincuenta y dos hisopos de muestras endocervicales de mujeres infértiles para detectar C. trachomatis. La identificación de los genotipos de C. trachomatis se realizó mediante la técnica de PCR basado en RFLP y en el método automatizado de secuenciación para el gen ompA. Para identificar el genotipo, la secuencia de 891 pb obtenida se alineó con secuencias del gen ompA de Chlamydia disponibles en el GenBank. Resultados Veinticuatro mujeres con infertilidad (15,8%) fueron positivas para C. trachomatis. De acuerdo con los métodos de RFLP y de secuencición de nucleótidos, el genotipo más frecuente correspondió al serotipo F (n=13 [54,2%]), seguida por el serotipo E (n=2 [8,7%]), G (n=2 [8,7%]), K (n=2 [8,7%]) y L2 (n=2 [8,7%]), mientras que los serotipos D, H e Ia fueron menos frecuentes (todos, n=1 [4,2%]). Ninguno de los pacientes con resultado positivo para el serotipo L2 tuvo síntomas de linfogranuloma venéreo (LGV). El análisis de las secuencias nucleotídicas del serotipo L2 mostró una nueva genovariante diferente de la L2b a la L2F. Los puntos de mutación en esta nueva genovariante se observaron en el dominio VS1 del gen ompA. Conclusiones En este estudio el genotipo más frecuente fue el F. Además, se demostró la presencia del genotipo L2 en mujeres infértiles que no mostraron signos y síntomas de la enfermedad de LGV. Se evidenciaron mutaciones puntuales en las secuencias nucleotídicas del genotipo L2 por lo cual hay necesidad de una mayor investigación para identificar la existencia de nuevas genovariantes L2 en América Latina (AU)

In vitro study of biosynthesis of protoporphyrin IX induced by Ù-Aminolevulinic acid in normal and cancerous cells of the human cervix

Bxkground. Ù-Aminolevulinic acid (ALA) is recognized as the starter in the biosynthesis of the heme group, the structural basis of cytochromes, chlorophylls, biliary pigments, and other porphyrins. It is the first intermediary in the biosynthesis of protoporphyrin IX (PpIX), and of ther heme group. PpIX is present in low concentration in normal cells, and in high concentration in tumor cells. Methods. The accumulation of protoporhyrin IX (PpIX) induced by Ù-aminolevulinic acid (ALA) was tested in two cervico-uterine cancer cell lines (HeLa and CaLo), and in normal human cervical epithelial (NHCE) cells. Results. The optimal concentration of ALA that induced maximum levels of intra- and extracellular accumulation of PpIX in both HeLa and NHCE cells was 300 µg of ALA/mL, and for CaLo cells, 150 µg/mL. The viability of HeLa, CaLo, and NHCE cells exposed to ALA measured 81, 98 and 84 percent, respectively. The optimale time for accumulation of PpIX, both intra- and extracellular, was 4 h for HeLa and NHCE cells and 5 h for CaLo cells per 24 h of expusure to optimal concentrations of ALA. After the maximum level of PpIX accumulation was reached, there was a gradual decrease until there was only a small quantity. A statistically significant difference (p <0.0001) was foundf in the accumulation of PpIX, depending on the concentrations of ALA used as well as between cervical cancer cell lines and 1:7, and for NHCE and CaLo cells. 1:5. Conclusions. These results are important for determining the usefulness of the sensitizer (PpIX)