Diseño de un Marcador Molecular para Identificar el Segmento VS4 el Gen omp1 de Chlamydia Trachomatis en Mujeres Embarazadas con Presión Arterial Alta Asociada a Preeclampsia

Abstract

Chlamydia trachomatis is a gram negative bacteria intracellular by force which causes severe conditions (STD) for the health of women, men and children, which in the majority of cases don't present symptoms making its detection difficult. In addition it causes different pathologies depending on the host and the site of infection. To counter this problematic it was proposed to design an efficient, specific, sensible and economic molecular marker to identify the VS4 segment of the gene omp1 of the MOMP protein of Chlamydia trachomatis in pregnant women with preeclampsia, living in a specific sector of Bogotá, Colombia, and identify the serovar present in this segment, which cause genital illnesses or infections. To succeed, bioinformatic programs and tools such as GenBank, PickPrimer and Blast of NCBI on one hand, and NetPrimer and Primer3 on the other hand, have been used to find the theoretical support for the idea and for design of these molecular markers, while for the amplification of the segment conventional chain reactions of polimerasa, in warm medium and in real time were used in order to, finally, read out the DNA segments using electrophoresis in an agarose gel. Three pairs of molecular markers were obtained, called SV2, SV4 and SV3 which amplified the segments of 128pb (region VS3), 277pb (region VS4) and 282pb (region VS3-VS4) respectively, in male and female using the HotStartPCR technique. Additionally, the marker pair SV3 showed a high sensibility and specifity as it already could identify the presence of C. trachomatis in DNA samples of only 3ng/µl doing the test with qPCR. As far as the serovar were concerned, ten (10) derivatives of the variants B, D, E, F, G and L, originators of genital-urinary and eye disorders were recognized. It can be concluded that the molecular markers designed in this investigation though they need to be applied with the HotStartPCR or qPCR, fulfill the awaited requirements and permit the detection of this STD in men as in women. However it is recommended to realize a study focused on pregnant women with preeclampsia for the simple reason that this part of the study could not be accomplished with.