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.

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.

Spores of Trichoderma Strains over P. vulgaris Beans: Direct Effect on Insect Attacks and Indirect Effect on Agronomic Parameters.

Acanthoscelides obtectus is an insect pest that attacks wild and cultivated common beans (Phaseolus vulgaris L). Four Trichoderma strains, the T. arundinaceum IBT 40837 wild-type strain (=Ta37), a producer of trichothecene harzianum A (HA), two transformants of T. arundinaceum strain, Ta37-17.139 (=Δtri17) and Ta37-23.74 (=Δtri23), and the T. brevicompactum IBT 40841 wild-type strain (=Tb41), which produces the trichothecene trichodermin, were assessed to establish their direct effect on insect attacks and their indirect effect on the plants grown from the beans treated with those fungal strains and exposed to insect attacks. Treatments of bean seeds with different Trichoderma strains led to different survival rates in the insects, and the Tb41 strain caused the lowest survival rate of all. An 86.10% of the insect cadavers (in contact with Δtri23) showed growth of this strain. This was the treatment that attracted the greatest number of insects. The daily emergence was reduced in beans treated with the Ta37, Tb41, and Δtri17 strains. The undamaged beans treated with Ta37 and Δtri23 showed a high capacity of germination (80.00% and 75.00%, respectively), whereas the Δtri17 and Tb41 treatments increased the capacity of germination in the damaged beans (66.67%). The undamaged beans treated with Δtri23 had the greatest dry weights for the aerial part (4.22 g) and root system in the plants (0.62 g). More studies on the mechanisms of insect control, plant growth promotion, and trichodermol and trichodermin production by Δtri23 and Tb41, respectively, should be explored in order to commercialize these fungal species on a large scale.

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.

Xylotrechus arvicola (Coleoptera: Cerambycidae) capture in vineyards in relation to climatic factors.

BACKGROUND: Captures and seasonal abundance of Xylotrechus arvicola (Coleoptera: Cerambycidae) in relation to climatic factors were studied in vineyards between the years 2013 and 2020. Insects captures from vine wood in two Vitis vinifera varieties were evaluated every year by counting the number of insects captured with CROSSTRAP®. The captured insects were grouped (by sex and total) into ranges of 10 days and compared to climatic data (daily average, temperature and rainfall) for each cultivar and year. RESULTS: The capture periods spanned from 1 June and 31 July, with the period from 1 to 30 June having the greatest number of insect captures, as long as weather conditions were favourable, i.e. temperature above 20.00 °C and accumulated rainfall in 10 days lower than 0.40 mm, verified through the analysis of parameter estimates, in which, only the temperature parameter was significantly. CONCLUSIONS: The study provided useful information for the integrated pest management of X. arvicola through mass trapping in vineyards when temperature exceeds 20.00 °C and the accumulated rainfall is less than 0.40 mm in 10 days to obtain peak captures. This is the first quantitative study of X. arvicola control associated with temperature and rainfall in Vitis vinifera. © 2022 Society of Chemical Industry.

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.

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.

Effect of Cry Toxins on Xylotrechus arvicola (Coleoptera: Cerambycidae) Larvae.

The beetle Xylotrechus arvicola is a destructive pest in vineyards (Vitis vinifera) in the main wine-producing areas of the Iberian Peninsula. X. arvicola larvae bore into the grapevine wood-making galleries, thus damaging the plant both directly and indirectly; the latter through the proliferation of wood fungi, which can invade the inside of the plant, decreasing the quality and quantity of its production. The susceptibility of X. arvicola larvae to five coleopteran toxic Cry proteins (Cry1B, Cry1I, Cry3A, Cry7A, and Cry23/37) was evaluated under laboratory conditions in order to deepen the knowledge of the effect of these proteins on this insect throughout its biological development. Cry7Ab and Cry1Ba were the most effective in controlling X. arvicola larvae due to the significant reduction in larvae survival (32.9 and 25.9 days, respectively), and by causing serious alterations in the larvae during the remaining months of their development. The developmental stage of the prepupal and pupal stages was not affected by the previous ingestion of Cry proteins. The Cry proteins tested could be applied to control X. arvicola larvae since they were able to kill them and cause serious alterations in the larvae during the remaining months of development that followed. The data presented suggest that these Cry proteins can be used as bioinsecticides against the larvae of this insect, applying them only at the moment when the larvae hatch from the egg outside the grapevine wood (this would only be useful and justified if the economic threshold is exceeded) in order to avoid the rapid evolution of resistance against these toxins since not all of the larvae were killed and thus increase vine wood protection.

[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.

Detección atípica de Chlamydia pneumoniae no humana en una muestra endocervical. Reporte de caso / Atypical detection of non-human Chlamydia pneumoniae in an endocervical sample. Case Report

Resumen ANTECEDENTES: Chlamydia trachomatis es uno de los principales microorganismos de trasmisión sexual asociado de manera importante con infertilidad femenina. La detección de genotipos y nuevas variantes de Chlamydia trachomatis permite conocer su prevalencia, distribución geográfica, identificar la aparición de resistencia antimicrobiana y las asociaciones clínicas o comportamientos sexuales y desarrollar vacunas. Este caso clínico es el primer informe de infección endocervical por una cepa diferente a C trachomatis. CASO CLÍNICO: Paciente de 25 años, con diagnóstico de infertilidad primaria de 2 años de evolución por factor endocrino-ovárico (sobrepeso e hipotiroidismo subclínico) y por factor masculino de hipospermia y teratozoospermia. El cultivo microbiológico endocervical detectó la infección por Ureaplasma spp y Chlamydia spp. La identificación de la cepa de Chlamydia mediante secuenciación del gen 16S del ARNr informó que era Chlamydia pneumoniae. La existencia de un plásmido en esta cepa de C pneumoniae confirmó que la infección endocervical fue por una cepa de Chlamydia pneumoniae no humana. CONCLUSIÓN: Este caso clínico sugiere la posibilidad de que una cepa de C pneumoniae no humana sea capaz de trasmitirse sexualmente a los humanos, estar circulando en la población mexicana y causar infertilidad, aunque aún se desconocen el origen y la dirección de la trasmisión.

Abstract BACKGROUND: Chlamydia trachomatis is one of the leading sexually transmitted microorganisms that is significantly associated with the development of female infertility. The detection of genotypes and new variants ofChlamydia trachomatisallows us to know their prevalence and geographic distribution, identify the appearance of antimicrobial resistance, clinical associations, or sexual behaviors, and develop vaccines. This clinical case reports for the first time endocervical infection by a strain other thanC. trachomatis. CLINICAL CASE: A 25-year-old woman with primary infertility of 2 years of evolution due to endocrine-ovarian factor (overweight and subclinical hypothyroidism) and male factor characterized by hypospermia and teratozoospermia. Endocervical microbiological culture detected infection byUreaplasma urealyticumandChlamydiaspp. Identification of theChlamydiastrain by sequencing the 16S rRNA gene reported that it wasChlamydia pneumoniae. The presence of plasmid in this strain ofC. pneumoniaeconfirmed that the endocervical infection was by a non-humanChlamydia pneumoniaestrain. CONCLUSION: This clinical case suggests that a non-human strain ofC. pneumoniaecan be sexually transmitted to humans, circulating in the Mexican population, and causing infertility, although the origin and direction of transmission are still unknown.

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.

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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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.

Misidentification of Burkholderia pseudomallei and Other Burkholderia Species From Pediatric Infections in Mexico.

Burkholderia pseudomallei and Burkholderia cepacia complex are poorly studied in Mexico. The genotypic analysis of 38 strains isolated from children with pneumonia were identified and showed that both Burkholderia groups were present in patients. From our results, it is plausible to suggest that new species are among the analyzed strains.

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.

[Prevalence of genital Chlamydia trachomatis infection in women attending in the National Institute of Perinatology from Mexico City]. / Prevalencia de la infección genital por Chlamydia trachomatis en mujeres que asisten al Instituto Nacional de Perinatología de la Ciudad de México.

BACKGROUND: Endocervical infection by Chlamydia trachomatis is considered one of the leading causes of infertility worldwide. During pregnancy, it can lead to serious complications such as premature rupture of membranes and premature births. AIM: To determine the prevalence of genital infection by C. trachomatis in pregnancy and infertile women from Mexico City. METHODS: The detection of C. trachomatis was performed by real-time PCR with the commercial kit COBAS® TaqMan CT Test v2.0 (Roche Molecular System). RESULTS: We analyzed 2,352 endocervical swabs; 102 were positive (4.3%). Age prevalence showed that pregnant adolescents (15 to 19 years of age) had the highest risk of infection (10.9%, RR = 3.23 [95% IC: 1.79-5.84]), followed by young women aged 20 to 24 years, with a prevalence of 5.6% (RR = 1.65 [95% IC: 0.82-3.34]). DISCUSSION: The results indicate that the prevalence is within the range reported worldwide. However, pregnant adolescents were those with a higher prevalence than infertile women were. CONCLUSION: A systematic screening of C. trachomatis infection in women younger than 24 years of age, and in pregnant women is necessary to reduce the incidence of infertility and perinatal complications.

Prevalencia de la infección genital por Chlamydia trachomatis en mujeres que asisten al Instituto Nacional de Perinatología de la Ciudad de México / Prevalence of genital Chlamydia trachomatis infection in women attending in the National Institute of Perinatology from Mexico City

Resumen Introducción: La infección endocervical por Chlamydia trachomatis es considerada una de las principales causas de infertilidad en todo el mundo. Durante el embarazo puede conducir a complicaciones graves como la ruptura prematura de membranas y los partos prematuros. Objetivo: Determinar la prevalencia de infección genital por C. trachomatis en mujeres embarazadas e infértiles de la Ciudad de México. Métodos: La detección de C. trachomatis fue mediante reacción de polimerasa en cadena tiempo real (RPC-TR) con el kit comercial COBAS® TaqMan CT Test v2,0 (Roche Molecular System). Resultados: Se analizaron 2.352 muestras; 102 fueron positivas (4,3%). La prevalencia por edad mostró que las adolescentes embarazadas (15 a 19 años) fueron las de mayor riesgo de infección (10,9%, RR = 3,23 [IC 95%: 1,79-5,84]), seguido de mujeres jóvenes de 20 a 24 años, con prevalencia de 5,6% (RR = 1,65 [IC 95%: 0,82-3,34]). Discusión: Los resultados indican que la prevalencia está dentro del rango reportado en el concierto mundial. Sin embargo, las adolescentes embarazadas tuvieron mayor prevalencia que las mujeres infértiles. Conclusión: Es imperioso realizar un rastreo sistemático de infección por C. trachomatis en mujeres bajo 24 años de edad, y en mujeres embarazadas para disminuir los casos de infertilidad y las complicaciones perinatales.

Background: Endocervical infection by Chlamydia trachomatis is considered one of the leading causes of infertility worldwide. During pregnancy, it can lead to serious complications such as premature rupture of membranes and premature births. Aim: To determine the prevalence of genital infection by C. trachomatis in pregnancy and infertile women from Mexico City. Methods: The detection of C. trachomatis was performed by real-time PCR with the commercial kit COBAS® TaqMan CT Test v2.0 (Roche Molecular System). Results: We analyzed 2,352 endocervical swabs; 102 were positive (4.3%). Age prevalence showed that pregnant adolescents (15 to 19 years of age) had the highest risk of infection (10.9%, RR = 3.23 [95% IC: 1.79-5.84]), followed by young women aged 20 to 24 years, with a prevalence of 5.6% (RR = 1.65 [95% IC: 0.82-3.34]). Discussion: The results indicate that the prevalence is within the range reported worldwide. However, pregnant adolescents were those with a higher prevalence than infertile women were. Conclusion: A systematic screening of C. trachomatis infection in women younger than 24 years of age, and in pregnant women is necessary to reduce the incidence of infertility and perinatal complications.