PCR múltiplex para el diagnóstico de microorganismos atípicos transmitidos sexualmente / Multiplex PCR testing for the atypical microorganisms diagnosis sexually transmitted

Resumen Antecedentes: Las infecciones de transmisión sexual son un problema de salud pública mundial. El análisis rutinario incluye solo pruebas microbiológicas y serológicas para el diagnóstico de patógenos. Los microorganismos atípicos como Chlamydia trachomatis y micoplasmas no son identificados debido a los requerimientos. Además, no es incluida Gardnerella vaginalis, aunque se asocia a la vaginosis bacteriana. Objetivo: Desarrollar una PCR múltiplex para el diagnóstico de C. trachomatis, micoplasmas y G. vaginalis. Método: Se estandarizó la PCR múltiplex utilizando oligonucleótidos para C. trachomatis (gen ompA, orf6 plasmídico), Mycoplasma/Ureaplasma y G. vaginalis (genes rRNA16s). Resultados: Se estandarizaron pruebas de PCR múltiplex para los microorganismos estudiados, optimizándose las concentraciones y condiciones de las reacciones múltiplex. Se obtuvieron PCR dúplex para C. trachomatis (ompA, orf6), Chlamydia/Gardnerella y Chlamydia/micoplasmas y tríplex para Chlamydia/Mycoplasma/Ureaplasma. También un cuádruplex para Chlamydia/Mycoplasma/Ureaplasma/Gardnerella. Los resultados fueron verificados por PCR e hibridación automática (HybriSpot 12) y análisis in silico. Conclusión: Se desarrollaron pruebas de PCR múltiplex con una alta sensibilidad y especificidad para la identificación de C. trachomatis, micoplasmas y G. vaginalis.

Abstract Background: Sexually transmitted infections are a global public health problem. Routine analysis includes microbiological and serological tests for the diagnosis of pathogens. Atypical microorganisms such as Chlamydia trachomatis and mycoplasmas are not determined due to the requirements for their identification. Furthermore, Gardnerella vaginalis is not included despite being associated with bacterial vaginosis. Objective: To develop a multiplex PCR to diagnose Chlamydia, mycoplasmas, and Gardnerella. Method: Standardization of multiplex PCR tests was carried out using oligonucleotides for the identification of Chlamydia (ompA gene, plasmid orf6), Mycoplasma/Ureaplasma and Gardnerella (rRNA16s genes). Results: Multiplex PCR tests were standardized for the microorganisms studied, optimizing the concentrations and conditions of the multiplex reactions. Duplex PCR was obtained for Chlamydia (ompA, orf6), Chlamydia/Gardnerella, and Chlamydia/mycoplasmas, and triplex PCR for Chlamydia/mycoplasmas. Also, a quadruplex for Chlamydia, Mycoplasma/Ureaplasma and Gardnerella. PCR and automatic hybridization verified the results obtained (HybriSpot 12) and in silico analysis. Conclusion: Multiplex PCR tests with high sensitivity and specificity were developed to identify C. trachomatis, mycoplasmas, and G. vaginalis.

Prevalencia de Chlamydia trachomatis en mujeres del Hospital General de Zona No. 29 / Chlamydia trachomatis prevalence in women from the Hospital General de Zona No. 29

Introducción: la Chlamydia trachomatis es la principal causa de infecciones bacterianas de transmisión sexual a nivel mundial. Se estima que cada año se producen 131 millones de casos. Cursa de manera asintomática, pero la infección ascendente en mujeres puede conducir a la enfermedad inflamatoria pélvica, embarazo ectópico e infertilidad. Objetivo: determinar la prevalencia de C. trachomatis en mujeres de población abierta que acuden al Hospital General de Zona No. 29. Material y métodos: se realizó la identificación de C. trachomatis por pruebas de PCR a 200 muestras de exudado vaginal y se determinó su genotipo. Paralelamente, se realizó el diagnóstico microbiológico de rutina. Resultados: la prevalencia de C. trachomatis fue del 8.5% (17/200) con una concomitancia significativa de p = 0.006 con Gardnerella vaginalis (riesgo relativo de 2.871, IC95%: 1.574-5.236). Asimismo, se identificó C. trachomatis en cinco muestras como el único agente etiológico. Dieciséis cepas de C. trachomatis pertenecieron al genotipo F. Una cepa identificada de C. trachomatis presentó motivos genéticos similares a la variante mexicana reportada en 2019. Conclusiones: la prevalencia de C. trachomatis en la población estudiada nos indica la necesidad de implementar técnicas de diagnóstico para esta bacteria. El uso de la PCR permite realizar una determinación genotípica rápida, que explicaría el comportamiento epidemiológico de la C. trachomatis y representaría una mejora significativa de la calidad de vida de la paciente.

Background: Chlamydia trachomatis is the main cause of sexually transmitted bacterial infections worldwide. An estimated of 131 million cases occur each year. It is asymptomatic, but ascending infection in women can lead to pelvic inflammatory disease, ectopic pregnancy, and infertility. Objective: To determine the prevalence of C. trachomatis in open population women who attend the Hospital General de Zona No. 29. Material and methods: Identification of C. trachomatis was carried out by PCR testing of 200 vaginal exudate samples and its genotype was determined. In parallel, a routine microbiological diagnosis was carried out. Results: The prevalence of C. trachomatis was 8.5% (17/200) with a significant concomitance of p = 0.006 with Gardnerella vaginalis (relative risk of 2.871, 95%CI: 1.574- 5.236). Likewise, C. trachomatis was identified in 5 samples as the only etiological agent. Sixteen strains of C. trachomatis belong to genotype F. An identified strain of C. trachomatis presented genetic motifs similar to the Mexican variant repor- ted in 2019. Conclusions: The prevalence of C. trachomatis in the studied population indicates the need to implement diagnostic techniques for this bacterium. The use of PCR allows a rapid genotypic determination that would explain the epidemiological behavior of C. trachomatis and would represent a sig- nificant improvement in the quality of life of the patient.

[Chlamydia trachomatis prevalence in women from the Hospital General de Zona No. 29]. / Prevalencia de Chlamydia trachomatis en mujeres del Hospital General de Zona No. 29.

BACKGROUND: Chlamydia trachomatis is the main cause of sexually transmitted bacterial infections worldwide. An estimated of 131 million cases occur each year. It is asymptomatic, but ascending infection in women can lead to pelvic inflammatory disease, ectopic pregnancy, and infertility. OBJECTIVE: To determine the prevalence of C. trachomatis in open population women who attend the Hospital General de Zona No. 29. MATERIAL AND METHODS: Identification of C. trachomatis was carried out by PCR testing of 200 vaginal exudate samples and its genotype was determined. In parallel, a routine microbiological diagnosis was carried out. RESULTS: The prevalence of C. trachomatis was 8.5% (17/200) with a significant concomitance of p = 0.006 with Gardnerella vaginalis (relative risk of 2.871, 95%CI: 1.574-5.236). Likewise, C. trachomatis was identified in 5 samples as the only etiological agent. Sixteen strains of C. trachomatis belong to genotype F. An identified strain of C. trachomatis presented genetic motifs similar to the Mexican variant reported in 2019. CONCLUSIONS: The prevalence of C. trachomatis in the studied population indicates the need to implement diagnostic techniques for this bacterium. The use of PCR allows a rapid genotypic determination that would explain the epidemiological behavior of C. trachomatis and would represent a significant improvement in the quality of life of the patient.

INTRODUCCIÓN: la Chlamydia trachomatis es la principal causa de infecciones bacterianas de transmisión sexual a nivel mundial. Se estima que cada año se producen 131 millones de casos. Cursa de manera asintomática, pero la infección ascendente en mujeres puede conducir a la enfermedad inflamatoria pélvica, embarazo ectópico e infertilidad. OBJETIVO: determinar la prevalencia de C. trachomatis en mujeres de población abierta que acuden al Hospital General de Zona No. 29. MATERIAL Y MÉTODOS: se realizó la identificación de C. trachomatis por pruebas de PCR a 200 muestras de exudado vaginal y se determinó su genotipo. Paralelamente, se realizó el diagnóstico microbiológico de rutina. RESULTADOS: la prevalencia de C. trachomatis fue del 8.5% (17/200) con una concomitancia significativa de p = 0.006 con Gardnerella vaginalis (riesgo relativo de 2.871, IC95%: 1.574-5.236). Asimismo, se identificó C. trachomatis en cinco muestras como el único agente etiológico. Dieciséis cepas de C. trachomatis pertenecieron al genotipo F. Una cepa identificada de C. trachomatis presentó motivos genéticos similares a la variante mexicana reportada en 2019. CONCLUSIONES: la prevalencia de C. trachomatis en la población estudiada nos indica la necesidad de implementar técnicas de diagnóstico para esta bacteria. El uso de la PCR permite realizar una determinación genotípica rápida, que explicaría el comportamiento epidemiológico de la C. trachomatis y representaría una mejora significativa de la calidad de vida de la paciente.

Prevalence, concordance and reproductive sequelae after Chlamydia trachomatis infection in Mexican infertile couples.

There are few concordance studies on the Chlamydia trachomatis (infection among infertile couples. The objective of this research was to know the prevalence, concordance and reproductive sequelae that couples may develop when both partners show a C. trachomatis infection. A cross-sectional study among 688 infertile couples using the C. trachomatis detection by real-time PCR was performed. The infertility causes were obtained from their medical records. The prevalence of infection was 8.68%. The percentage of concordance was 22.4% (13 couples). A presence of tubal occlusion was only associated with infected-discordant women [RR = 3.46, 95% CI (1.54-7.74), p < .003]. Seminal values were not associated with discordant men. The concordant couples showed association with the infection and tubal occlusion [RR = 3.19, 95% CI (1.09-9.34), p < .05], and oligozoospermia [RR = 12.17, 95% CI (4.29-34.54), p < .001], hypospermia [RR = 14.13, 95% CI (4.78-41.84), p < .001]. An alteration in semen quality was shown particularly in men whose sexual partners show a tubal pathology. This could occur due to a C. trachomatis infection in the testis, which underlines the need to carry out effective and efficient strategies to identify and treat all sexual partners exposed to C. trachomatis.

Use of the mtrR Gene for Rapid Molecular Diagnosis of Neisseria gonorrhoeae and Identification of the Reduction of Susceptibility to Antibiotics in Endocervical Swabs.

BACKGROUND: Neisseria gonorrhoeae is one of the main etiological agents of sexually transmitted diseases. The asymptomatic course of the infection and its resistance to antibiotics can lead to pelvic inflammatory disease and infertility. OBJECTIVES: We developed a polymerase chain reaction (PCR) test using the methyltetrahydrofolate homocysteine methyltransferase reductase (mtrR) gene to identify N. gonorrhoeae and detect reduced susceptibility to antibiotics. MATERIAL AND METHODS: We analysed 250 samples of endocervical exudate from infertile women with a negative diagnosis of N. gonorrhoeae. We designed NGmtr primers to detect N. gonorrhoeae and identify the antibiotic-resistant strain. RESULTS: Of the 250 samples, 60 (24%) tested positive for N. gonorrhoeae using real-time PCR. Our study was validated using the HO primers and the Seeplex STD6 ACE System, with a 100% correlation. Furthermore, the NGmtr primers are specific for N. gonorrhoeae and not for other species. Additionally, the curves generated by real-time PCR differed between wild and variant strains (10.93%). The dissociation temperatures for the wild and variant strains were 86.5 and 89 °C, respectively. CONCLUSIONS: The NGmtr primers enabled us to identify N. gonorrhoeae strains with or without reduction of susceptibility to antibiotics. Therefore, this work constitutes a tool that will facilitate the diagnosis of this infection for a low cost and improve patient quality of life.

Estimación de la carga de enfermedad por cáncer de cuello uterino en Ecuador / Estimation of the burden of disease for cervical cancer in Ecuador

El cáncer de cuello uterino es la segunda causa de muerte por cáncer en mujeres en Ecuador.[1] Pese a la disponibilidad de intervenciones de prevención primaria como la inmunización contra virus de papiloma humano y de diferentes estrategias de tamizaje mediante citología y/o pruebas moleculares, las tasas de mortalidad por cáncer de cuello uterino en la mayoría de países de América Latina se mantienen altas.[2] Después de evaluar su alta costo-efectividad, USD$ 597 por cada año de vida ajustado por discapacidad (AVAD) evitado desde la perspectiva de la sociedad, el Ministerio de Salud Pública del Ecuador introdujo el año 2014 la inmunización contra el virus del papiloma humano (VPH) en su programa ampliado de inmunizaciones.[3] También se ha evaluado que el uso de pruebas moleculares de detección de tipos de VPH de alto riesgo es altamente costo-efectiva como estrategia de tamizaje, con US$ 3482 por años vividos con discapacidad (AVAD) evitado desde la perspectiva de la sociedad.[4] Si bien la introducción de la inmunización contra VPH es promisoria en términos de reducir la carga de enfermedad asociada a cáncer de cuello uterino, sus resultados serán visibles a largo plazo, por lo que es imprescindible el fortalecimiento de los programas de tamizaje y tratamiento oportuno, y la monitorización de la efectividad de estas intervenciones

Cervical cancer is the second cause of cancer death in women in Ecuador. [1] Despite the availability of interventions by primary prevention such as immunization against papilloma virus and different screening strategies using cytology and / or Molecular tests, death rates from cervical cancer in most Latin American countries they remain high. [2] After evaluating its high cost-effectiveness, USD $ 597 per each disability-adjusted life year (DALY) prevented from the perspective of society, the Ministry of Public Health of Ecuador introduced the year 2014 the immunization against the papilloma virus Human (HPV) in its expanded program of immunizations. [3] As well it has been evaluated that the use of molecular tests to detect High-risk HPV types are highly cost-effective as a strategy of screening, with US $ 3482 for years lived with disability (DALY) avoided from the perspective of society. [4] While the introduction of immunization against HPV is promising in terms of reducing the burden of disease associated with cervical cancer, its results will be visible in the long term, which is why the strengthening of screening programs and timely treatment, and monitoring the effectiveness of these interventions

La definición del perfil profesional del médico familiar en Ecuador: una tarea en proceso / The definition of the professional profile of the family doctor in Ecuador: a task in progress

El funcionamiento adecuado de los modelos de salud familiar y comunitaria propuestos tanto por el Ministerio de Salud Pública como por el instituto Ecuatoriano de Seguridad Social, dependerá en gran medida de la incorporación de recursos humanos altamente capacitados en la estrategia de Atención Primaria en Salud, entre los que destaca el rol de los médicos especialistas en medicina familiar y comunitaria. En Ecuador la formación de especialistas en medicina familiar inició en 1987, con la implementación del primer programa de Especialidad en Medicina Familiar y permitió la fundación de la Sociedad Ecuatoriana de Medicina Familiar un año después. Tras 23 años de labor de los diferentes programas de especialización en medicina familiar, contamos con una masa crítica de más de dos centenares de médicos de familia formados en Ecuador, decididos a afrontar el desafío de reperflar e incrementar exponencialmente el número de especialistas en atención primaria requeridos en el país. Presentamos un resumen del trabajo iniciado en noviembre del 2008 por la Sociedad Ecuatoriana de Medicina Familiar para redefnir el perfl del médico familiar en Ecuador, y que debe considerarse como una tarea en proceso. El borrador de esta propuesta concluye que el médico de familia es un profesional sanitario especializado en proporcionar atención de salud integral, continuada y de calidad a todo individuo en el contexto de su familia y a las familias en el contexto de su comunidad y los determinantes de la salud; utiliza el enfoque y la estrategia de atención primaria en salud, con el fn de promover, mantener e incrementar el bienestar de las personas en congruencia con los valores e ideales de la sociedad y de los conocimientos profesionales actuales.

The proper functioning of family and community health models proposed by both the Ministry of Public Health and the Ecuadorian Institute of Social Security, will depend largely on the incorporation of human resources highly trained in the strategy of Primary Health Care, among which highlights the role of medical specialists in family and community medicine. In Ecuador, the training of specialists in family medicine began in 1987, with the implementation of the first program of Specialty in Family Medicine and allowed the foundation of the Ecuadorian Society of Family Medicine a year later. After 23 years of work of the different programs of specialization in family medicine, we have a critical mass of more than two hundred family doctors trained in Ecuador, determined to face the challenge of impacting and exponentially increasing the number of primary care specialists required in the country. We present a summary of the work initiated in November 2008 by the Society Ecuadorian Family Medicine to redefine the profile of the family doctor in Ecuador, and that should be considered as a work in progress. The draft of this proposal concludes that the family doctor is a healthcare professional specialized in providing comprehensive, continuous and quality health care to every individual in the context of your family and families in the context of your community and the determinants of health; uses the primary health care approach and strategy, with in order to promote, maintain and increase the well-being of people in congruence with the values ​​and ideals of society and current professional knowledge.

Hipoparatiroidismo postquirúrgico y calcificaciones cerebrales extensas en en hombre de 64 años / Post-surgical hypoparathyroidism and extensive brain calcifications in a 64-year-old man

Reportamos el caso de un hombre de 64 años admitido en el servicio de emergencias por convulsiones. La tomografía computada cerebral reveló calcificaciones simétricas extensas de los hemisferios cerebrales, tálamo,ganglios de la base y cerebelo, El reporte radiológico sugirió enfermedad de Fahr, un trastorno neurodegenerativo con depósitos de calcio característicos en ganglios de la base y otras áreas cerebrales visibles en estudios de neuroimagen. Las calcificaciones cerebrales simétricas identificadas radiográficamente se presentan en una gran variedad de condiciones. Nuestro paciente fue sometido a una tireidectomía hace 46 años, que postoperatoriamente resultó en hipotirodismo e hipoparatiroidismo. Este es el primer caso reportado en nuestro país de hipoparatiroidismo posquirúrgico con calcificaciones cerebrales externsas.

We report the case of a 64-year-old man admitted to the emergency service for seizures. Cerebral computed tomography revealed extensive symmetrical calcifications of the cerebral hemispheres, thalamus, base ganglia, and cerebellum. The radiological report suggested Fahr's disease, a neurodegenerative disorder with characteristic calcium deposits in base ganglia, and other brain areas visible in studies. neuroimaging. Radiographically identified symmetric brain calcifications occur under a wide variety of conditions. Our patient underwent a thyroidectomy 46 years ago, which postoperatively resulted in hypothyroidism and hypoparathyroidism. This is the first reported case of postsurgical hypoparathyroidism in our country with external brain calcifications.

[Immune response of suckling mouse brain (CRL) rabies vaccine and tissue culture rabies vaccine (Verorab) in pre-exposure prophylaxis in humans]. / Evaluación de la capacidad inmunogénica de la vacuna antirrábica tipo Fuenzalida- Palacios (CRL) y de la vacuna antirrábica de cultivo celular (Verorab) en personas con tratamiento preexposición.

BACKGROUND: A WHO experts committee recommended the substitution of antirabic vaccines produced in nervous tissue, by vaccines produced in tissue cultures. AIM: To compare the immunogenic capacity of antirabic vaccines CRL (produced in nervous tissue) and Verorab (produced in tissue culture), used for pre-exposure prophylaxis in humans. PATIENTS AND METHODS: Fifty four volunteers were immunized for this study. The first group, vaccinated with CLR was treated with a scheme of 4 subcutaneous peri umbilical doses in days 0, 3, 7 and 28. The second group, vaccinated with Verorab vaccine was treated with a scheme of 3 intramuscular doses in deltoid zone at days 0, 7 and 28. Blood samples were obtained at days 0, 7, 42 and 365 to measure neutralizing antibodies using the Inhibition of Fluorescent Focus Technique (RFFIT). RESULTS: At day seven, a primary non protective immunologic response was observed in both groups, with titers significantly higher in the group vaccinated with Verorab. At day 42, no differences were observed. At day 365, all subjects vaccinated with Verorab and 50% of individuals vaccinated with CRL had protective antibody titers (p <0.05). CONCLUSIONS: The vaccine produced in cell breeds (Verorab) produces a better an faster immunologic response compared to the CRL vaccine.