Frecuencia y detección molecular de Gardnerella vaginalis en una institución de tercer nivel / Frequency and molecular detection of Gardnerella vaginalis in a third level institute

Resumen Antecedentes: Los padecimientos vaginales son la razón más común para que las mujeres busquen atención médica, con una prevalencia global que oscila entre el 23 y el 29% en mujeres en edad reproductiva. La vaginosis bacteriana es una de las principales causas de estos padecimientos, y el agente etiológico más frecuentemente identificado es Gardnerella vaginalis, sin embargo su diagnóstico es difícil, ya que requiere de medios artificiales selectivos enriquecidos y diferenciales. Objetivo: Determinar la frecuencia de G. vaginalis mediante la amplificación de ácidos nucleicos empleando la reacción en cadena de la polimerasa (PCR) en muestras cervicovaginales de pacientes que asisten a un instituto de tercer nivel. Método: Se analizaron 121 muestras cervicovaginales para la detección molecular del ARN ribosomal 16S de G. vaginalis. Resultados: G. vaginalis. se detectó en 34 muestras, de estas, 23 fueron de mujeres embarazadas y 11 de no embarazadas. Conclusión: La PCR de punto final detectó tres veces más la presencia de G. vaginalis que el medio de cultivo artificial.

Abstract Background: Vaginal conditions are the most common reason for women to seek medical care, with an overall prevalence ranging from 23 to 29% in women of reproductive age. Bacterial vaginosis is one of the main causes of these conditions, and the most frequently identified etiological agent is Gardnerella vaginalis, however, its diagnosis is difficult since it requires enriched and artificial selective culture media. Objective: To determine the frequency of G. vaginalis by nucleic acid amplification using polymerase chain reaction (PCR) in cervicovaginal samples from patients attending a third level institute. Method: One hundred twenty-one cervicovaginal samples were analyzed for molecular detection of 16S ribosomal RNA from G. vaginalis. Results: G. vaginalis was detected in 34 samples, of these, 23 were from pregnant women and 11 from non-pregnant women. Conclusion: Endpoint PCR detected three times more the presence of G. vaginalis than artificial culture medium.

Análisis molecular y filogenético del genotipo F de Chlamydia trachomatis aislados en México en 2011 / Molecular and phylogenetic analysis of the F genotype of Chlamydia trachomatis isolated in Mexico in 2011

Resumen Antecedentes: Chlamydia trachomatis es la bacteria que se detecta con mayor frecuencia en las infecciones de transmisión sexual. Se han identificado 20 genotipos de C. trachomatis mediante el gen ompA y varias genovariantes mediante el análisis de polimorfismo de un solo nucleótido (SNP). En México, el genotipo F es el más frecuente. Objetivo: Identificar la existencia de subtipos del genotipo F. Método: Se analizaron siete cepas del genotipo F de C. trachomatis aisladas en 2011, mediante secuenciación de nucleótidos y mapeo con enzimas de restricción. Resultados: El análisis de SNP mostró dos cepas con el mismo SNP en el nucleótido 288 (C288T), mientras que con enzimas de restricción se identificó una variante con diferente RFLP (polimorfismo de la longitud de fragmentos de restricción) cuando se tratan con la mezcla de enzimas HinfI y TaqI. Conclusión: En México se encuentran dos subtipos del genotipo F y solo las enzimas de restricción HinfI y TaqI pueden identificar la existencia de uno de estos genotipos F.

Abstract Background: Chlamydia trachomatis is the most frequently identified bacterium in sexually transmitted infections. Twenty C. trachomatis genotypes have been determined using the ompA gene and several genovariants by single nucleotide polymorphism (SNP) analysis. In Mexico, the F genotype is the most frequent. Objective: To identify subtypes of the F genotype. Method: Seven C. trachomatis genotype F strains isolated in 2011 were analyzed by nucleotide sequencing and restriction enzyme mapping. Results: SNP analysis showed two strains with the same SNP at nucleotide 288 (C288T), while with res-triction enzymes, a variant with different RFLP (restriction fragment length polymorphism) was identified when treated with the mixture of HinfI and TaqI enzymes. Conclusion: In Mexico, there are two subtypes of F, and only with restriction enzymes HinfI and TaqI can identify one of the genovariants of the F genotype.

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.

Modelling the Unidentified Abortion Burden from Four Infectious Pathogenic Microorganisms (Leptospira interrogans, Brucella abortus, Brucella ovis, and Chlamydia abortus) in Ewes Based on Artificial Neural Networks Approach: The Epidemiological Basis for a Control Policy.

Unidentified abortion, of which leptospirosis, brucellosis, and ovine enzootic abortion are important factors, is the main cause of disease spread between animals and humans in all agricultural systems in most developing countries. Although there are well-defined risk factors for these diseases, these characteristics do not represent the prevalence of the disease in different regions. This study predicts the unidentified abortion burden from multi-microorganisms in ewes based on an artificial neural networks approach and the GLM. METHODS: A two-stage cluster survey design was conducted to estimate the seroprevalence of abortifacient microorganisms and to identify putative factors of infectious abortion. RESULTS: The overall seroprevalence of Brucella was 70.7%, while Leptospira spp. was 55.2%, C. abortus was 21.9%, and B. ovis was 7.4%. Serological detection with four abortion-causing microorganisms was determined only in 0.87% of sheep sampled. The best GLM is integrated via serological detection of serovar Hardjo and Brucella ovis in animals of the slopes with elevation between 2600 and 2800 meters above sea level from the municipality of Xalatlaco. Other covariates included in the GLM, such as the sheep pen built with materials of metal grids and untreated wood, dirt and concrete floors, bed of straw, and the well water supply were also remained independently associated with infectious abortion. Approximately 80% of those respondents did not wear gloves or masks to prevent the transmission of the abortifacient zoonotic microorganisms. CONCLUSIONS: Sensitizing stakeholders on good agricultural practices could improve public health surveillance. Further studies on the effect of animal-human transmission in such a setting is worthwhile to further support the One Health initiative.

Identificación de genotipos de Chlamydia trachomatis en neonatos con distrés respiratorio / Identification of Chlamydia trachomatis genotypes in newborns with respiratory distress

Introducción: Cada año se notifican ciento treinta millones de infecciones por Chlamydia trachomatis en todo el mundo. Diecinueve serotipos de este patógeno pueden causar infecciones en mujeres embarazadas y recién nacidos. En México se desconoce la distribución de estos genotipos en recién nacidos con infecciones respiratorias. Material y métodos: Se analizaron mil sesenta y dos muestras de lavado bronquial de neonatos con síndrome de dificultad respiratoria para detección de infección por clamidia. El diagnóstico de clamidia se realizó mediante la detección de plásmidos con un ensayo PCR interno y los genotipos se identificaron mediante un ensayo PCR-RFLP del gen ompA. Resultados: El genotipado de 40 cepas identificó a 14 como I/Ia (35%), 13 como E (32,5%), 7 como D (17,5%), 5 como F (12,5%) y 1 como L2 (2,5%). El análisis de riesgo relativo mostró que el genotipo D se asoció con sepsis neonatal (RR=5,83; IC 95%: 1,51-25,985; p <0,02), mientras que el genotipo I/Ia mostró asociación significativa con madres que desarrollaron corioamnionitis (2,8; IC 95%: 1,4-5,5; p <0,05). Conclusiones: Si bien los genotipos I/Ia y E de Chlamydia trachomatis fueron la causa más frecuente de infección respiratoria en neonatos mexicanos, el 80% de los genotipos F produjeron este padecimiento. En cambio, el genotipo D se asoció con el desarrollo de sepsis neonatal y el genotipo I/Ia con corioamnionitis. (AU)

Introduction: One hundred thirty million Chlamydia trachomatis infections are reported worldwide each year. Nineteen serotypes of this pathogen can cause infection in pregnant women and neonates. The distribution of these genotypes in newborns with respiratory infections in Mexico is unknown. Material and methods: We tested 1062 bronchial lavage samples from neonates with respiratory distress syndrome for Chlamydia infection. The diagnosis of Chlamydia was made by plasmid detection with an in-house PCR assay, and genotypes were identified using a PCR-RFLP assay for the ompA gene. Results: The genotyping of 40 strains identified 14 as I/Ia (35%), 13 as E (32.5%), 7 as D (17.5%), 5 as F (12.5%), and 1 as L2 (2.5%). The relative risk analysis showed that genotype D was associated with neonatal sepsis (RR, 5.83; 95% confidence interval [CI], 1.51-25.985; P<.02), while the I/Ia genotype was significantly associated with chorioamnionitis in the mother (2.8; 95% CI, 1.4–5.5; P<.05). Conclusions: Although Chlamydia trachomatis genotypes I/Ia and E of were the strains involved most frequently in respiratory infections in Mexican neonates, 80% of patients with genotype F developed respiratory disease. In contrast, genotype D was associated with neonatal sepsis, and genotype I/Ia with chorioamnionitis. (AU)

Identification of Chlamydia trachomatis genotypes in newborns with respiratory distress.

INTRODUCTION: One hundred thirty million Chlamydia trachomatis infections are reported worldwide each year. Nineteen serotypes of this pathogen can cause infection in pregnant women and neonates. The distribution of these genotypes in newborns with respiratory infections in Mexico is unknown. MATERIAL AND METHODS: We tested 1062 bronchial lavage samples from neonates with respiratory distress syndrome for Chlamydia infection. The diagnosis of Chlamydia was made by plasmid detection with an in-house PCR assay, and genotypes were identified using a PCR-RFLP assay for the ompA gene. RESULTS: The genotyping of 40 strains identified 14 as I/Ia (35%), 13 as E (32.5%), 7 as D (17.5%), 5 as F (12.5%), and 1 as L2 (2.5%). The relative risk analysis showed that genotype D was associated with neonatal sepsis (RR, 5.83; 95% confidence interval [CI], 1.51-25.985; P < .02), while the I/Ia genotype was significantly associated with chorioamnionitis in the mother (2.8; 95% CI, 1.4-5.5; P < .05). CONCLUSIONS: Although C. trachomatis genotypes I/Ia and E of were the strains involved most frequently in respiratory infections in Mexican neonates, 80% of patients with genotype F developed respiratory disease. In contrast, genotype D was associated with neonatal sepsis, and genotype I/Ia with chorioamnionitis.

Evaluation of the goat cellular immune response to rBtuB-Hia-FlgK peptides from Brucella melitensis.

Brucellosis is a zoonosis caused by Brucella; B. melitensis is the most prevalent species in goats and humans. Previously, three B. melitensis peptides, rBtuB-Hia-FlgK showed antigen-specific immune responses in rodent models. The goal of this study was to evaluate the goat Th1/Th2 immune response to B. melitensis peptides. Twenty-eight animals were separated into four groups and were immunized with the rBtuB-Hia-FlgK peptides cocktail, adjuvant, PBS and Rev-1 vaccine, respectively. Peripheral blood samples were collected on days 0, 15, and 80 post-inoculation. The CD4+ and CD8+ T cells proliferation, and cytokine production of the Th-1 (IL-2, IL-12, TNF-α, and IFN-γ) and Th-2 profiles (IL-4, IL-5, and IL-10) were evaluated. An increase of CD4+/CD8+ at 15 days post-vaccination was observed and continued until the 80th. In addition, the IFN-γ, TNF-α, and IL-2 mRNA expression were typically induced by the 15th day, but only IFN-γ levels were observed at day 80 post-immunization. Brucella pathogenesis is distinguished by the presence of a large amount of Th-1 cytokines. Although a reduced amount of IFN-γ in the culture supernatant was accurately detected compared with Rev-1 after 15 days, it could be influenced by the sampling schedule, as a higher cytokine production might be induced as early as the first-week post-vaccination. The results indicate that rBtuB-Hia-FlgK induced an immune response similar to the Rev-1 vaccine. The possible use of inert molecules with the unique ability to typically induce cellular response similar to attenuated vaccine represents an attractive option that should not be ruled out.

Identification of Chlamydia trachomatis genotypes in Mexican men with infertile women as sexual partners / Identificación de genotipos de Chlamydia trachomatis en hombres mexicanos con mujeres infértiles como parejas sexuales

Background: Chlamydia trachomatis is considered a public health problem due to the high prevalence in sexually active women and men. The distribution of genital Chlamydia genotypes among Mexican men is unknown. Objective: To assess the prevalence of Chlamydia genotypes in men with infertile women as sexual partners. Methods: A total of 659 urine samples were collected from men whose sexual partners were infertile women; the identifying Chlamydia infection was by means of a real-time nucleic acid amplification test (qPCR). OmpA gene PCR-RFLP and sequencing were used to confirm the genotypes of C. trachomatis. The association of genotypes with age, spermatic parameters and gynecological data of sexual partners was further analyzed. Results: Forty-nine urine samples were positive infection (7.4%). The Chlamydia infection was significantly associated with teratozoospermia, azoospermia, hypospermia, and oligozoospermia. Five genotypes (F 51%; 12.2% to D; 12.2% to E; 6.1% to L2 and 4.1% Ia) were correctly identified. None genotypes identified in this comparative study were positively associated with changes in some of the spermatic values because all of them typically produce some considerable damage to these cells. Conclusions: The F genotype was the most frequent genotype identified in infertile men from Mexico City and all genotypes play an important role in the seminal alteration of Mexican men whose female partners are infertile.(AU)

Antecedentes: Chlamydia trachomatis se considera un problema de salud pública debido a la alta prevalencia en mujeres y hombres sexualmente activos. Se desconoce la distribución de los genotipos genitales de Chlamydia entre los hombres mexicanos.Objetivo: Evaluar la prevalencia de los genotipos de Chlamydia en hombres con mujeres infértiles como parejas sexuales. Métodos: Se recogieron 659 muestras de orina de hombres cuyas parejas sexuales eran mujeres infértiles; la identificación de la infección por Chlamydia se realizó mediante una prueba de amplificación de ácido nucleico en tiempo real (qPCR). Se utilizaron la PCR-RFLP y la secuenciación del gen OmpA para confirmar los genotipos de C. trachomatis. Se analizó en mayor profundidad la asociación de los genotipos con la edad, los parámetros espermáticos y los datos ginecológicos de las parejas sexuales. Resultados: Cuarenta y nueve muestras de orina dieron positivo para la infección (7,4 %). La infección por Chlamydia se asoció significativamente con la teratozoospermia, la azoospermia, la hipospermia y la oligozoospermia. Se identificaron correctamente cinco genotipos (F 51 %; 12,2 % para D; 12,2 % para E; 6,1 % para L2 y 4,1 % Ia). Ninguno de los genotipos identificados en este estudio comparativo se asoció positivamente con cambios en algunos de los valores espermáticos porque todos ellos suelen producir algún daño considerable en estas células. Conclusiones: El genotipo F fue el más frecuente identificado en hombres infértiles de Ciudad de México y todos los genotipos desempeñan un papel importante en la alteración seminal de los hombres mexicanos cuyas parejas femeninas son infértiles.(AU)

Identification of Chlamydia trachomatis genotypes in Mexican men with infertile women as sexual partners.

BACKGROUND: Chlamydia trachomatis is considered a public health problem due to the high prevalence in sexually active women and men. The distribution of genital Chlamydia genotypes among Mexican men is unknown. OBJECTIVE: To assess the prevalence of Chlamydia genotypes in men with infertile women as sexual partners. METHODS: A total of 659 urine samples were collected from men whose sexual partners were infertile women; the identifying Chlamydia infection was by means of a real-time nucleic acid amplification test (qPCR). OmpA gene PCR-RFLP and sequencing were used to confirm the genotypes of C. trachomatis. The association of genotypes with age, spermatic parameters and gynecological data of sexual partners was further analyzed. RESULTS: Forty-nine urine samples were positive infection (7.4%). The Chlamydia infection was significantly associated with teratozoospermia, azoospermia, hypospermia, and oligozoospermia. Five genotypes (F 51%; 12.2% to D; 12.2% to E; 6.1% to L2 and 4.1% Ia) were correctly identified. None genotypes identified in this comparative study were positively associated with changes in some of the spermatic values because all of them typically produce some considerable damage to these cells. CONCLUSIONS: The F genotype was the most frequent genotype identified in infertile men from Mexico City and all genotypes play an important role in the seminal alteration of Mexican men whose female partners are infertile.

Identification of Chlamydia trachomatis genotypes in Mexican men with infertile women as sexual partners.

BACKGROUND: Chlamydia trachomatis is considered a public health problem due to the high prevalence in sexually active women and men. The distribution of genital Chlamydia genotypes among Mexican men is unknown. OBJECTIVE: To assess the prevalence of Chlamydia genotypes in men with infertile women as sexual partners. METHODS: A total of 659 urine samples were collected from men whose sexual partners were infertile women; the identifying Chlamydia infection was by means of a real-time nucleic acid amplification test (qPCR). OmpA gene PCR-RFLP and sequencing were used to confirm the genotypes of C. trachomatis. The association of genotypes with age, spermatic parameters and gynecological data of sexual partners was further analyzed. RESULTS: Forty-nine urine samples were positive infection (7.4%). The Chlamydia infection was significantly associated with teratozoospermia, azoospermia, hypospermia, and oligozoospermia. Five genotypes (F 51%; 12.2% to D; 12.2% to E; 6.1% to L2 and 4.1% Ia) were correctly identified. None genotypes identified in this comparative study were positively associated with changes in some of the spermatic values because all of them typically produce some considerable damage to these cells. CONCLUSIONS: The F genotype was the most frequent genotype identified in infertile men from Mexico City and all genotypes play an important role in the seminal alteration of Mexican men whose female partners are infertile.

Co-infection between genotypes of the human papillomavirus and Chlamydia trachomatis in Mexican women.

Not all human papillomavirus (HPV) infections develop into cervical cancer (CC), so it is proposed that other factors may influence this, such as co-infection with Chlamydia trachomatis (CT). To identify the prevalence of co-infection, we included 189 women with suspicion of HPV. Viral typing was performed by carrying out the Roche HP Linear Array test, while CT detection was performed with the COBAS® TaqMan® 48 kit from Roche. Of the 189 women only 184 had an infection with HPV, CT or both: 56.6% were positive for one or several HPV genotypes, and 67.7% for CT. Clinical data showed an association between HPV and CIN I (n = 22; RR = 2.43; 95% CI 1.72-3.43, p < 0.05). CT infection was only associated with cervicitis (n = 40; RR = 1.73; 95% CI 1.34-2.23, p < 0.05). The CT-HPV co-infection rate was 28%. Co-infection revealed an association with CIN I (n = 31, RR= 3.33; 95% CI 2.08-5.34 p < 0.05), CIN III (n = 7; RR = 2.57; 95% CI 1.53-4.31, p < 0.05); and a significant risk of 2.3 (95% CI 1.08-4.90) times higher to develop CC; nevertheless, this risk was not statistically significant. CT/HPV co-infection was associated with the development of a high-grade lesion (CIN III) as well as an important risk for developing CC.

Riesgo de infección en recién nacidos expuestos a la colonización vaginal por Chlamydia trachomatis / Risk of infection in newborns exposed to vaginal colonisation by Chlamydia trachomatis

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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.

Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis

Chlamydia trachomatis is considered as a public health problem due to its high prevalence and increased rates of gynecological disorders. The major outer membrane protein (MOMP) of this bacterium is the most abundant protein in its membrane and has been evaluated not only as a vaccine development candidate but also is used in many diagnostic tests. The MOMP weighs 69 kDa and contains four variable segments (VS 1-4) separated by constant regions. Several research groups have developed recombinant single-variable segments of MOMP expressed in Escherichia coli cytoplasm. But, all variable segments have been used minimally for the diagnosis of a chlamydial infection. In this experiment, the authors obtained the recombinant MOMP of C. trachomatis (rMOMP) in E. coli rMOMP and extracted, purified, and partially characterized it. This was later used to identify anti-Chlamydia trachomatis antibodies in sera of infertile patients by immunodetection assays, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence tests. The ELISA test showed high sensitivity and low specificity of 100 and 58.3%, respectively. The above results obtained were linked to the cross-reactivity of antibodies against C. pneumoniae or C. psittaci. Hence, an evaluation was performed to obtain an optimized test for the diagnosis of C. trachomatis infection

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A randomized controlled trial comparing isosorbide dinitrate-oxytocin versus misoprostol-oxytocin at management of foetal intrauterine death.

BACKGROUND: The metabolic activity of endogenous nitric oxide (NO) and the medical use of nitrovasodilatory drugs like isosorbide dinitrate have been shown to be potential inducers inducers of cervical ripening prior to surgical evacuation of the uterus. OBJECTIVE: To assess the therapeutic efficacy and safety of combined isosorbide dinitrate-oxytocin in the management of intrauterine foetal death (IUFD). METHODS: Sixty women with IUFD after 20 weeks of gestation requesting uterine evacuation were randomly selected to receive isosorbide dinitrate gel solution (80 mg/1.5 mL; n = 30) or misoprostol gel solution (100 mcg/1.5 mL; n = 30) every 3 h with a maximum of four doses or until a Bishop score >7 was reached. Subsequently, patients received a high dose of intravenous oxytocin until complete uterus evacuation was achieved. Therapeutic efficacy was evaluated by mean the relative risk of the foetal expulsion based on comparison of event rates, and the proportion of women induced to labor at 7, 10 and 15 h after the administration of isosorbide dinitrate or misoprostol. Safety was assessed on the basis of woman´s vital signs and evaluation of adverse effects, including headache, abdominal pain, pelvic pain, lower back pain, nausea, dizziness and vomiting. RESULTS: The foetal expulsion rate using the isosorbide dinitrate-oxytocin combination was approximately 4.4 times, and at least 2.1 times, the foetal expulsion rate with the misoprostol-oxytocin regimen at any given point in time. The proportion of women achieved vaginal delivery at 15 hours was 100% for the isosorbide dinitrate-oxytocin group and 86.7% for the misoprostol-oxytocin group. The average delivery induction interval was significantly lower when isosorbide dinitrate-oxytocin was used (8.7 ± 3.1 h) than when misoprostol-oxytocin (11.9 ± 3.1 h) was used. A total of 20% of patients in the isosorbide dinitrate-oxytocin group recorded headache, and no cases of uterine tachysystole, haemorrhage or coagulopathy were recorded. CONCLUSION: This study indicates that intravaginal isosorbide dinitrate followed by intravenous oxytocin was more effective than the conventional method used to induce labour in the medical management of foetal death in pregnancies after 20 weeks of gestation. TRIAL REGISTRATION: Clinicaltrials.gov NCT02488642.

Obtaining an ELISA test based on a recombinant protein of Chlamydia trachomatis.

Chlamydia trachomatis is considered as a public health problem due to its high prevalence and increased rates of gynecological disorders. The major outer membrane protein (MOMP) of this bacterium is the most abundant protein in its membrane and has been evaluated not only as a vaccine development candidate but also is used in many diagnostic tests. The MOMP weighs 69 kDa and contains four variable segments (VS 1-4) separated by constant regions. Several research groups have developed recombinant single-variable segments of MOMP expressed in Escherichia coli cytoplasm. But, all variable segments have been used minimally for the diagnosis of a chlamydial infection. In this experiment, the authors obtained the recombinant MOMP of C. trachomatis (rMOMP) in E. coli rMOMP and extracted, purified, and partially characterized it. This was later used to identify anti-Chlamydia trachomatis antibodies in sera of infertile patients by immunodetection assays, enzyme-linked immunosorbent assay (ELISA), and indirect immunofluorescence tests. The ELISA test showed high sensitivity and low specificity of 100 and 58.3%, respectively. The above results obtained were linked to the cross-reactivity of antibodies against C. pneumoniae or C. psittaci. Hence, an evaluation was performed to obtain an optimized test for the diagnosis of C. trachomatis infection.

Identification of a new variant of Chlamydia trachomatis in Mexico / Identificación de nueva variante de Chlamydia trachomatis en México

Introduction: Chlamydia trachomatis is one of the main etiological agents of sexually transmitted infections worldwide. In 2006, a Swedish variant of C. trachomatis (Swedish-nvCT), which has a deletion of 377bp in the plasmid, was reported. In Latin America, Swedish-nvCT infections have not been reported. We investigated the presence of Swedish-nvCT in women with infertility in Mexico. Methods: Swedish-nvCT was searched in 69 C. trachomatis positive samples from 2339 endocervical specimens. We designed PCR primers to identify the deletion in the plasmid in the ORF1, and the presence of a repeated 44 bp in the ORF3. The sample with the deletion was genotyped with the genes of the major outer membrane protein A (ompA) and the polymorphic membrane protein (pmpH). Results: The deletion was detected in one of the 69 samples positive C. trachomatis of 2339 endocervical exudates. The nucleotide sequence analysis of the ompA shows a high degree of similarity with the Swedish nvCT (98%), however the variant found belongs to serovar D. The nucleotide sequence of the pmpH gene associates to the variant found in the genitourinary pathotype of the Swedish-nvCT but in different clusters. Conclusions: Our results revealed the presence of a new variant of C. trachomatis in Mexican patients. This variant found in Mexico belongs to serovar D based on the in silico analysis of the ompA and pmpH genes and differs to the Swedish-nvCT (serovars E). For these variants of C. trachomatis that have been found it is necessary to carry out a more detailed analysis, although the role of this mutation has not been demonstrated in the pathogenesis

Introducción: Chlamydia trachomatis es una de las principales bacterias que causan infecciones de transmisión sexual en todo el mundo. En 2006 se informó de una variante sueca de C. trachomatis (nvCT-sueca), que tiene una deleción de 377 bp en su plásmido. En América Latina no se ha informado de infecciones por la nvCT-sueca. El propósito de esta investigación fue la búsqueda de la nvCT-sueca en mujeres mexicanas con infertilidad. Métodos: Se analizaron 69 muestras positivas para C. trachomatis de 2.339 muestras endocervicales. Se diseñaron cebadores que identificaron la deleción de 377 pb en ORF1, y detección de un tándem de 44 pb repetidos en ORF3, como ocurre en la nvCT-sueca. Las muestras con la deleción fueron genotipificadas mediante los genes de la proteína principal de la membrana externa A (ompA) y de la proteína polimórfica de membrana H (pmpH). Resultados: La deleción se detectó en una de las 69 muestras (1,44%). El análisis de la secuencia del gen ompA mostró un alto grado de similitud con la nvCT-sueca (98%). Sin embargo, la variante encontrada perteneció al serovar D. La secuencia del gen pmpH se asoció al patotipo genitourinario, pero en diferentes clusters al de la nvCT-sueca. Conclusiones: Los resultados revelaron la presencia de una nueva variante de C. trachomatis en México con delección y que pertenece al serovar D con base al análisis in silico de los genes ompA y pmpH, y que difiere de la nvCT-sueca (serovares E). Se requiere conocer su prevalencia en México y en América Latina

Identification of a new variant of Chlamydia trachomatis in Mexico.

INTRODUCTION: Chlamydia trachomatis is one of the main etiological agents of sexually transmitted infections worldwide. In 2006, a Swedish variant of C. trachomatis (Swedish-nvCT), which has a deletion of 377bp in the plasmid, was reported. In Latin America, Swedish-nvCT infections have not been reported. We investigated the presence of Swedish-nvCT in women with infertility in Mexico. METHODS: Swedish-nvCT was searched in 69C. trachomatis positive samples from 2339 endocervical specimens. We designed PCR primers to identify the deletion in the plasmid in the ORF1, and the presence of a repeated 44bp in the ORF3. The sample with the deletion was genotyped with the genes of the major outer membrane protein A (ompA) and the polymorphic membrane protein (pmpH). RESULTS: The deletion was detected in one of the 69 samples positive C. trachomatis of 2339 endocervical exudates. The nucleotide sequence analysis of the ompA shows a high degree of similarity with the Swedish nvCT (98%), however the variant found belongs to serovar D. The nucleotide sequence of the pmpH gene associates to the variant found in the genitourinary pathotype of the Swedish-nvCT but in different clusters. CONCLUSIONS: Our results revealed the presence of a new variant of C. trachomatis in Mexican patients. This variant found in Mexico belongs to serovar D based on the in silico analysis of the ompA and pmpH genes and differs to the Swedish-nvCT (serovars E). For these variants of C. trachomatis that have been found it is necessary to carry out a more detailed analysis, although the role of this mutation has not been demonstrated in the pathogenesis.