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1.
Article in English | MEDLINE | ID: mdl-30970111

ABSTRACT

Multidrug resistance (MDR), virulence and transferable elements potentiate Pseudomonas aeruginosa's role as an opportunistic pathogen creating a high risk for public health. In this study, we evaluated the possible association of multidrug resistance, virulence factors and integrons with intrahospital P. aeruginosa strains isolated from patients at Cumana hospital, Venezuela. Relevant clinical-epidemiological data were collected to study 176 strains (2009-2016) isolated from different hospital units. Bacterial resistance was classified as susceptible, low-level resistant (LDR), multidrug resistant (MDR) and extensively drug-resistant (XDR). Most strains produced pyoverdine, DNase, gelatinase and hemolysin. Around 73% of the strains showed some type of movement. MDR and XDR strains increased from 2009 (24.2% and 4.8%, respectively) to 2016 (53.1% and 18.8%); while LDR decreased from 64.5% to 6.3%. The exoU and exoS genes were found in a significant number of strains (38.1 and 7.4%, respectively). Class I integrons were detected in 35.8% of the strains and the frequency was associated with resistance (42.9, 22.4, 41.4 and 61.9%, for susceptible, LDR, MDR and XDR, respectively). The MDR/XDR strains were positively associated with hemolysins and exoU, but negatively associated with bacterial twitching. MDR/XDR phenotypes were also associated with the Intensive Care Unit (ICU), septicemia, bronchial infection and diabetic foot ulcers, as well as long hospital stay (≥10 days) and previous antimicrobial treatment. High frequency of MDR/XDR strains and their association with class I integrons and virulence factors can increase the infection potential, as well as morbidity and mortality of patients attending this hospital and could spread infection to the community, creating a health risk for the region.


Subject(s)
Anti-Bacterial Agents/pharmacology , Integrons/genetics , Pseudomonas Infections/microbiology , Pseudomonas aeruginosa/pathogenicity , Virulence Factors/genetics , Biofilms/growth & development , Drug Resistance, Multiple, Bacterial , Hospitals, University , Humans , Intensive Care Units , Microbial Sensitivity Tests , Phenotype , Pseudomonas aeruginosa/drug effects , Pseudomonas aeruginosa/enzymology , Venezuela , Virulence
2.
Biomédica (Bogotá) ; 36(supl.1): 118-127, dic. 2016. graf, tab
Article in Spanish | LILACS | ID: lil-783529

ABSTRACT

Introducción. Escherichia coli es uno de los principales agentes causales del síndrome diarreico agudo. Objetivo. Identificar grupos clonales de E. coli enteropatógena en 485 casos de diarrea aguda en niños entre 0 y 10 años de edad atendidos en centros de salud de los municipios de Arismendi, Benítez y Sucre del estado Sucre, Venezuela, entre marzo y diciembre de 2011. Materiales y métodos. Previo consentimiento informado, se recolectaron muestras fecales y se identificó E. coli mediante coprocultivo estándar y serología con antisueros polivalentes y monovalentes. Se aisló el ADN y se amplificaron los genes eae (intimina) y bfpA (bundlina) mediante dos pruebas de reacción en cadena de la polimerasa (PCR) múltiples. Resultados. En 39,6 % de los coprocultivos se determinó la presencia de infección bacteriana. La prevalencia de E. coli fue de 54,7 %; 82,9 % de estas cepas fue positivo por serología para los serogrupos y el serotipo evaluados, principalmente en niños entre los 0 y los 2 años (37,9 %). El 48,6 % de las cepas de E. coli amplificaron para el gen eae y, de estas, 58,8 % se clasificó como cepas de E. coli enteropatógena típica (eae+ y bfp+). El ECEP II fue el serogrupo más frecuente (38,7 %), con predominio de bacterias E. coli enteropatógenas típicas (60 %). El alelo ß de la intimina fue el más identificado (74,5 %) en las cepas positivas para el gen eae. Solo se identificaron cuatro cepas con el serotipo O157:H7 utilizando antisueros, las cuales no amplificaron mediante PCR para los genes eae y bfpA. Conclusiones. Este estudio demostró la importancia de aplicar pruebas moleculares en la identificación de las cepas de E. coli causantes de diarrea de diversa gravedad.


Introduction: Diarrheagenic Escherichia coli is an important causative agent of acute diarrheic syndrome. Objective: To identify clonal groups of enteropathogenic E. coli (EPEC), in 485 children with acute diarrhea aged 0 to 10 years attending health care centers in Arismendi, Benítez and Sucre municipalities, Sucre state, Venezuela, from March to December, 2011. Materials and methods: After obtaining the informed consent, stool samples were collected. Escherichia coli was identified using standard coproculture methods and serology with polyvalent and monovalent antisera. DNA was isolated, and eae (intimin) and bfpA (bundlin) genes were amplified through two multiplex polymerase chain reactions (PCR). Results: The presence of bacterial infection was determined in 39.6% of coprocultures. The prevalence of E. coli was 54.7%; 82.9% of these isolates were positive by serology for the evaluated serogroups and serotypes, which were mostly identified in children between 0 and 2 years (37.9%); 48.6% of E. coli strains amplified the eae gene; of these, 58.8% were classified as typical EPEC (eae+ y bfp+). EPEC II was the most common serogroup (38.7%), with predominance of typical EPEC (60%). In positive strains for eae gene, the ß intimin allele was the most frequently identified (74.5%). Only four strains with O157:H7 serotype were identified, which showed no PCR amplification of the eae and bfpA genes. Conclusion: This study showed the importance of molecular tests to identify diarrheagenic E. coli strains causing clinical conditions of varying severity.


Subject(s)
Enteropathogenic Escherichia coli , Diarrhea , Escherichia coli , Gastrointestinal Diseases , Polymerase Chain Reaction
3.
Biomedica ; 36(0): 118-27, 2016 Mar 28.
Article in English | MEDLINE | ID: mdl-27622632

ABSTRACT

INTRODUCTION: Diarrheagenic Escherichia coli is an important causative agent of acute diarrheic syndrome.  OBJECTIVE: To identify clonal groups of enteropathogenic E. coli (EPEC), in 485 children with acute diarrhea aged 0 to 10 years attending health care centers in Arismendi, Benítez and Sucre municipalities, Sucre state, Venezuela, from March to December, 2011.  MATERIALS AND METHODS: After obtaining the informed consent, stool samples were collected. Escherichia coli was identified using standard coproculture methods and serology with polyvalent and monovalent antisera. DNA was isolated, and eae (intimin) and bfpA (bundlin) genes were amplified through two multiplex polymerase chain reactions (PCR).  RESULTS: The presence of bacterial infection was determined in 39.6% of coprocultures. The prevalence of E. coli was 54.7%; 82.9% of these isolates were positive by serology for the evaluated serogroups and serotypes, which were mostly identified in children between 0 and 2 years (37.9%); 48.6% of E. coli strains amplified the eae gene; of these, 58.8% were classified as typical EPEC (eae+ y bfp+). EPEC II was the most common serogroup (38.7%), with predominance of typical EPEC (60%). In positive strains for eae gene, the ß intimin allele was the most frequently identified (74.5%). Only four strains with O157:H7 serotype were identified, which showed no PCR amplification of the eae and bfpA genes.  CONCLUSION: This study showed the importance of molecular tests to identify diarrheagenic E. coli strains causing clinical conditions of varying severity.


Subject(s)
Diarrhea/epidemiology , Enteropathogenic Escherichia coli/isolation & purification , Escherichia coli Infections/epidemiology , Escherichia coli/isolation & purification , Polymerase Chain Reaction/methods , Child , Enteropathogenic Escherichia coli/chemistry , Escherichia coli/chemistry , Humans , Venezuela
4.
Ecancermedicalscience ; 9: 579, 2015.
Article in English | MEDLINE | ID: mdl-26557877

ABSTRACT

Cervical lesions have been associated with infection by high-risk human papilloma virus (high-risk HPV). In 409 women aged >15 years high-risk HPV lesions were identified. In a cohort of this population persistent infection was compared with cytological, colposcopic, and histological lesions. Cervical scrapes were taken and DNA was isolated. HPV was detected by PCR in the E6/E7 region. Genotyping was performed by PCR nested multiple E6/E7. HPV was detected in a 37.40% (153/409), high-risk HPV in 86% (153/178), HPV18 46.64% (83/178), HPV16 34.28% (61/178). Among these 53.93% (96/178) were multiple infections, and HPV18/16 (30/96) was the most frequent 31.25%. The cytology showed changes in 15% of positive patients. A 49.67% in women positive for HPV infection showed abnormalities in the colposcopic study, a relationship that turned out to be statistically significant ( p < 0.0019 test χ(2)). Among all 85% of the women were younger than 45 years of age. Fifty-seven patients were evaluated 15 months after the base study, with initial prevalence of morbidity 49.12% (28/57) and at the end 10.53% (6/57), showing in 89.29% (25/28) negative for HR-HPV infection, 10.34% (3/28) showed persistence of infection, 17.54% (10/57) presented cytological alterations, with 80% of positivity for HPV, and a regression of 100% (10/10) of the previously identified lesions. With colposcopy, 50% (14/28) presented alterations related to HPV, of these 85.71% (12/14) showed regression of such an alteration. The cumulative incidence for HPV was 10.34% (3/29). The incidence rate was 4.23% (3/71), which is equal to 4.23 new cases of HPV infection per 100 people, per year of follow-up. In conclusion, the present work shows a high frequency of infection by high-risk HPV, with predominance of HPV18 and 16 and in general for multiple infections. Colposcopy was better predictor than the Pap smear for infection. The follow-up study revealed a low percentage of persistent infection, and a high frequency of negativity for viral infection, high regression of cytological and colposcopic lesions, a low cumulative and incidence rate similar to that reported by other Latin American countries and higher than the European countries.

5.
Rev. Soc. Venez. Microbiol ; 34(2): 97-102, dic. 2014. tab
Article in Spanish | LILACS | ID: lil-746318

ABSTRACT

Un total de 1.203 individuos de cuatro poblaciones rurales y dos zonas urbanas de Cumaná, estado Sucre, Venezuela, se incluyeron en un estudio comparativo de parasitosis intestinales. Previa información y consentimiento se recolectaron muestras fecales que fueron procesadas por examen coproparasitológico, método de Kato cualitativo, Ritchie, tinción de Zielh Neelsen y cultivo en agar. El 77,8% de los individuos resultaron parasitados, hallándose diferencias entre éstos con el tipo de población evaluada (χ2=75,1; p<0,001). En las zonas rurales predominó el poliparasitismo y en las urbanas el monoparasitismo, hallándose diferencias significativas (χ2=136,1; p<0,001). En ambas zonas fue mas frecuente en el sexo femenino. La edad arrojó asociación significativa, según el tipo de población siendo más prevalente el parasitismo en los niños de 0 a7 años en zonas rurales y 8 a14 años en urbanas (χ2=22,6; p<0,004). Se diagnosticaron más especies en las zonas rurales siendo Blastocystis spp. (protozoario) y Trichuris trichiura (helminto) las de mayor prevalencia. Blastocystis spp. estuvo asociado con otros protozoarios. Los helmintos asociados fueron T. trichiura y Ascaris lumbricoides en los dos tipos de poblaciones. La alta frecuencia de parasitosis intestinales en las poblaciones evaluadas, demostró la exposición de los habitantes a mecanismos comunes de contaminación.


Abstract: We carried out a comparative study of intestinal parasites in 1.200 individuals from four rural and two urban populations located at Cumana, Sucre State, Venezuela. After previously obtaining their informed consent, we collected fecal samples that were processed by a coproparasitological examination, qualitative Kato’s method, Ritchie, Ziehl-Neelsen stain, and agar culture. Results showed that 77.8% of the individuals were parasitized, and that there were differences according to the type of population being evaluated (χ2=136.1; p<0.001). In both rural an urban areas predominated polyparasitism and monoparasitism respectively, finding significant differences (χ2=136.1; p<0.001). In both areas it was more frequent in females. Age showed a significant association and parasites were most prevalent in children 0 to 7 years old in rural areas and 8 to 14 years old in urban areas (χ2=22.6; p<0.004). More species were diagnosed in rural areas and Blastocytiis spp. (protozoa) and Trichuris trichuira (helminth) were the most prevalent. Blastocyttis spp. was associated with other protozoa. Associated helminths were T. trichuria and Ascaris lumbricoides in both types of populations. The high frequency of intestinal parasites in the populations evaluated shows the exposure of the inhabitants to common contamination mechanisms.

6.
Invest. clín ; 54(4): 392-405, dic. 2013. ilus, tab
Article in English | LILACS | ID: lil-740356

ABSTRACT

Genotyping of human papillomavirus (HPV) by molecular methods may enhance assessment information for screening and following of cervical infection. In this study, cervical samples were obtained from 250 women, along with colposcopic and cytological evaluations. A Nested-PCR-Multiplex assay was used for HPV detection and genotyping for HPV E6/E7 early regions. Infection with HPV was detected in 26.0% of the samples, with 98.46% positive for at least one genotype. High-risk HPVs were identified in 98.44%. HPV18 infection was detected in 76.92% of samples and HPV16 in 36.92%, whether as individual or as multiple infections. These infections were seen more frequently in women under 35 years of age (64.7%). The Pap-smear examination showed that 16.92% (11/65) of the samples had cervical changes suggesting HPV infection, whereas the colposcopic evaluation was suggestive of HPV infection in 47.69% (31/65) of DNA-HPV positive samples. There was a high frequency of high-risk HPV genotypes, particularly HPV18, alone or in multiple-type infections. Colposcopy findings showed to have a high predictive value for the diagnosis of HPV infection. The results reflect that over 50% of HPV-positive patients had a normal colposcopy and/or cytology, highlighting the importance of including HPV testing along with genotype identification in routine gynecological evaluations.


La genotipificación de virus del papiloma humano (VPH) por métodos moleculares puede proveer información valiosa para el monitoreo y seguimiento de la infección cervical. Se estudiaron muestras cervicales obtenidas a partir de 250 mujeres, en quienes se realizó, simultáneamente, evaluación citológica y colposcópica. Un ensayo de PCR, en formato Nested-múltiple para las amplificación de la región temparana E6/E7, fue realizado para la detección y genotipificación viral. La infección por VPH se detectó en 26,0% de las muestras, de las cuales, un 98,46% fue positivo para al menos uno de los genotipos probados; los VPH de alto riesgo se identificaron en un 98,44%. Los genotipos más frecuentes fueron VPH18 con un 76,92%, y VPH16 con un 36,92%, ya sea como infecciones individuales o múltiples. En cuanto a la edad, estas infecciones fueron más frecuentes en mujeres menores de 35 años, con un 64,7%. Los resultados citológicos mostraron que 16,92% (11/65) de las muestras cervicales tenían cambios sugestivos de infección por VPH; mientras que la evaluación colposcópica fue sugestiva de la infección por VPH en 47,69% (31/65) de las muestras ADN-VPH positivas. Se determinó una elevada frecuencia de los genotipos de VPH de alto riesgo, particularmente VPH18, solo o en infecciones múltiples. Los hallazgos colposcópicos mostraron un elevado valor predictivo para el diagnóstico de la infección por VPH. Los resultados reflejan que más del 50% de las pacientes VPH positivas tenían colposcopía y/o citología normal, evidenciando la importancia de incluir las pruebas de detección e identificación de VPH en la evaluación ginecológica de rutina.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Middle Aged , Young Adult , Nucleic Acid Amplification Techniques , Papillomaviridae/genetics , Genes, Immediate-Early , Genotype , Papillomaviridae/classification , Papillomavirus Infections/virology , Venezuela
7.
Invest Clin ; 54(4): 392-405, 2013 Dec.
Article in English | MEDLINE | ID: mdl-24502181

ABSTRACT

Genotyping of human papillomavirus (HPV) by molecular methods may enhance assessment information for screening and following of cervical infection. In this study, cervical samples were obtained from 250 women, along with colposcopic and cytological evaluations. A Nested-PCR-Multiplex assay was used for HPV detection and genotyping for HPV E6/E7 early regions. Infection with HPV was detected in 26.0% of the samples, with 98.46% positive for at least one genotype. High-risk HPVs were identified in 98.44%. HPV18 infection was detected in 76.92% of samples and HPV16 in 36.92%, whether as individual or as multiple infections. These infections were seen more frequently in women under 35 years of age (64.7%). The Pap-smear examination showed that 16.92% (11/65) of the samples had cervical changes suggesting HPV infection, whereas the colposcopic evaluation was suggestive of HPV infection in 47.69% (31/65) of DNA-HPV positive samples. There was a high frequency of high-risk HPV genotypes, particularly HPV18, alone or in multiple-type infections. Colposcopy findings showed to have a high predictive value for the diagnosis of HPV infection. The results reflect that over 50% of HPV-positive patients had a normal colposcopy and/or cytology, highlighting the importance of including HPV testing along with genotype identification in routine gynecological evaluations.


Subject(s)
Nucleic Acid Amplification Techniques , Papillomaviridae/genetics , Adolescent , Adult , Aged , Female , Genes, Immediate-Early , Genotype , Humans , Middle Aged , Papillomaviridae/classification , Papillomavirus Infections/virology , Venezuela , Young Adult
8.
Rev Soc Bras Med Trop ; 45(3): 329-33, 2012 Jun.
Article in English | MEDLINE | ID: mdl-22760131

ABSTRACT

INTRODUCTION: In Venezuela, acute diarrheic syndrome (ADS) is a primary cause of morbi-mortality, often involving the Salmonella genus. Salmonella infections are associated with acute gastroenteritis, one of the most common alimentary intoxications, and caused by the consumption of contaminated water and food, especially meat. METHODS: Conventional and molecular methods were used to detect Salmonella strains from 330 fecal samples from individuals of different ages and both sexes with ADS. Polymerase chain reaction (PCR) was used for the molecular characterization of Salmonella, using invA, sefA, and fliC genes for the identification of this genus and the serotypes Enteritidis and Typhimurium, respectively. RESULTS: The highest frequency of individuals with ADS was found in children 0-2 years old (39.4%), and the overall frequency of positive coprocultures was 76.9%. A total of 14 (4.2%) strains were biochemically and immunologically identified as Salmonella enterica subsp. enterica, of which 7 were classified as belonging to the Enteritidis serotype, 4 to the Typhimurium serotype, and 3 to other serotypes. The S. enterica strains were distributed more frequently in the age groups 3-4 and 9-10 years old. CONCLUSIONS: The molecular characterization method used proved to be highly specific for the typing of S. enterica strains using DNA extracted from both the isolated colonies and selective enrichment broths directly inoculated with fecal samples, thus representing a complementary tool for the detection and identification of ADS-causing bacteria.


Subject(s)
Diarrhea/microbiology , Gastroenteritis/microbiology , Salmonella Infections/microbiology , Salmonella/genetics , Acute Disease , Adolescent , Adult , Child , Child, Preschool , DNA, Bacterial/analysis , Feces/microbiology , Female , Gastroenteritis/diagnosis , Humans , Infant , Infant, Newborn , Male , Middle Aged , Polymerase Chain Reaction , Salmonella/classification , Salmonella/isolation & purification , Salmonella Infections/diagnosis , Sensitivity and Specificity , Serotyping , Venezuela , Young Adult
9.
Rev. Soc. Bras. Med. Trop ; 45(3): 329-333, May-June 2012. ilus, tab
Article in English | LILACS | ID: lil-640430

ABSTRACT

INTRODUCTION:In Venezuela, acute diarrheic syndrome (ADS) is a primary cause of morbi-mortality, often involving the Salmonella genus. Salmonella infections are associated with acute gastroenteritis, one of the most common alimentary intoxications, and caused by the consumption of contaminated water and food, especially meat. METHODS: Conventional and molecular methods were used to detect Salmonella strains from 330 fecal samples from individuals of different ages and both sexes with ADS. Polymerase chain reaction (PCR) was used for the molecular characterization of Salmonella, using invA, sefA, and fliC genes for the identification of this genus and the serotypes Enteritidis and Typhimurium, respectively. RESULTS: The highest frequency of individuals with ADS was found in children 0-2 years old (39.4%), and the overall frequency of positive coprocultures was 76.9%. A total of 14 (4.2%) strains were biochemically and immunologically identified as Salmonella enterica subsp. enterica, of which 7 were classified as belonging to the Enteritidis serotype, 4 to the Typhimurium serotype, and 3 to other serotypes. The S. enterica strains were distributed more frequently in the age groups 3-4 and 9-10 years old. CONCLUSIONS: The molecular characterization method used proved to be highly specific for the typing of S. enterica strains using DNA extracted from both the isolated colonies and selective enrichment broths directly inoculated with fecal samples, thus representing a complementary tool for the detection and identification of ADS-causing bacteria.


INTRODUÇÃO: Na Venezuela, síndrome da diarreia aguda (SDA) é a principal causa de mórbi-mortalidade, muitas vezes envolvem o gênero Salmonella. Infecções por Salmonella são associadas com gastroenterite aguda, uma das mais comuns intoxicações alimentares causada pelo consumo de água e alimentos contaminados, principalmente carne. MÉTODOS: Métodos convencionais e moleculares foram usados para detectar cepas de Salmonella em 330 amostras de fezes de indivíduos com SDA de diferentes idades e ambos os sexos. A reação em cadeia da polimerase (PCR) foi utilizada para a caracterização molecular de genes Salmonella invA, sefA e fliC para identificar o gênero e os sorotipos Enteritidis e Typhimurium, respectivamente. RESULTADOS: A maior frequência de indivíduos com SDA foi encontrada em crianças de 0-2 (39,4%) anos, e a frequência total de culturas de fezes positiva foi de 76,9%. Um total de 14 (4,2%) cepas foram bioquímica e imunologicamente identificados como Salmonella enterica subsp. enterica, dos quais 7 foram classificados como pertencentes ao sorotipo Enteritidis, Typhimurium sorotipo 4 e 3 para outros sorotipos. Cepas S. enterica foram distribuídas mais frequentemente em grupos de 3-4 e 9-10 anos de idade. CONCLUSÕES: O método de caracterização molecular usada provou ser altamente específico para tipificar as estirpes dos S. enterica usando tanto DNA extraído de colônias isoladas e direta e caldos de enriquecimento seletivo inoculados com amostras fecais, o que representa uma ferramenta complementar para a detecção e identificação de bactérias que causam a SDA.


Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Infant , Infant, Newborn , Male , Middle Aged , Young Adult , Diarrhea/microbiology , Gastroenteritis/microbiology , Salmonella Infections/microbiology , Salmonella/genetics , Acute Disease , DNA, Bacterial/analysis , Feces/microbiology , Gastroenteritis/diagnosis , Polymerase Chain Reaction , Sensitivity and Specificity , Serotyping , Salmonella Infections/diagnosis , Salmonella/classification , Salmonella/isolation & purification , Venezuela
10.
Invest. clín ; 52(4): 344-357, dic. 2011. ilus, tab
Article in English | LILACS | ID: lil-659224

ABSTRACT

High risk HPV infection is considered to play a central role in cervical carcinogenesis. HPV DNA testing has shown to be a very useful tool for screening and following cervical infections. The aim of this study was to compare three methods for HPV DNA detection, along with cytology and colposcopy analysis. Cervical samples were collected from 100 sexually active women in Mérida, western Venezuela. HPV infection was screened using Hybrid-Capture 2 (HC2), L1-Nested-PCR and E6/E7-PCR assays. 40% of the samples (40/100) were HPV positive by at least one of the DNA detection methods. HC2 detected HPV in 12% specimens. L1- and E6/E7-PCRs showed 50% sensitivity and 77% specificity.The agreement rate between HC2 and both PCR assays was 65%. Kappa value showed moderate concordance between HC2 and both PCR methods (κ=0.55; CI 95%). Also moderate concordance was seen when L1- and E6/E7-PCRs were compared (κ=0.48; CI 95%). There was a significant association between the Schiller test and E6/E7-PCR (p=0.006) for HPV infection. An acceptable agreement between all three assays for HPV detection was observed. Nevertheless, different PCR formats need to be further analyzed in order to make the right choice of method for HPV testing.


La infección con VPH de alto riesgo es el principal factor etiológico asociado al desarrollo de carcinogénesis cervical y las pruebas de detección de ADN-VPH han mostrado ser una herramienta esencial para la pesquisa y seguimiento de estas infecciones. El objetivo del estudio ha sido comparar tres métodos para la detección del ADN viral, en combinación con los análisis colposcópico y citológico. Se obtuvieron muestras cervicales de 100 mujeres sexualmente activas, en Mérida, Venezuela. La detección de infecciones por VPH se realizó por Captura Híbrida 2 (CH2) y los ensayos de PCR “L1-Nested-PCR” y “E6/E7-PCR”. 40% de las muestras (40/100) fueron positivas para VPH por al menos uno de los métodos aplicados. 12% de las muestras analizadas fueron positivas para VPH por CH2. Las dos PCR utilizadas mostraron un 50% de sensibilidad y 77% de especificidad. La coincidencia observada entre CH2 y las dos PCR fue del 65%. La determinación del valor Kappa mostró una concordancia moderada entre CH2 y ambos métodos de PCR (κ=0,55; CI 95%). También existió concordancia moderada al comparar las PCR de las regiones L1 y E6/E7 de VPH (κ=0,48; CI 95%). Hubo una asociación significativa entre el resultado del test de Schiller y la PCR E6/E7 (p=0,006) para la infección por VPH. Se determinó una concordancia aceptable entre los tres métodos aplicados para la detección de VPH; sin embargo, las PCR deben ser analizadas en trabajos futuros con el fin de establecer las pruebas más adecuadas para la detección viral.


Subject(s)
Adolescent , Adult , Aged , Female , Humans , Middle Aged , Alphapapillomavirus/isolation & purification , Cervix Uteri/virology , DNA Probes, HPV , DNA, Viral/analysis , Polymerase Chain Reaction/methods , Vaginal Smears , Alphapapillomavirus/genetics , Colposcopy , Consensus Sequence , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/virology , Genome, Viral , Nucleic Acid Amplification Techniques/methods , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/pathology , Papillomavirus Infections/virology , Reproducibility of Results , Sensitivity and Specificity , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/virology , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/virology , Uterine Cervicitis/pathology , Uterine Cervicitis/virology
11.
Invest Clin ; 52(4): 344-57, 2011 Dec.
Article in English | MEDLINE | ID: mdl-22523844

ABSTRACT

High risk HPV infection is considered to play a central role in cervical carcinogenesis. HPV DNA testing has shown to be a very useful tool for screening and following cervical infections. The aim of this study was to compare three methods for HPV DNA detection, along with cytology and colposcopy analysis. Cervical samples were collected from 100 sexually active women in Mérida, western Venezuela. HPV infection was screened using Hybrid-Capture 2 (HC2), L1-Nested-PCR and E6/E7-PCR assays. 40% of the samples (40/100) were HPV positive by at least one of the DNA detection methods. HC2 detected HPV in 12% specimens. L1- and E6/E7-PCRs showed 50% sensitivity and 77% specificity.The agreement rate between HC2 and both PCR assays was 65%. Kappa value showed moderate concordance between HC2 and both PCR methods (kappa=0.55; CI 95%). Also moderate concordance was seen when L1- and E6/E7-PCRs were compared (kappa=0.48; CI 95%). There was a significant association between the Schiller test and E6/E7-PCR (p=0.006) for HPV infection. An acceptable agreement between all three assays for HPV detection was observed. Nevertheless, different PCR formats need to be further analyzed in order to make the right choice of method for HPV testing.


Subject(s)
Alphapapillomavirus/isolation & purification , Cervix Uteri/virology , DNA Probes, HPV , DNA, Viral/analysis , Polymerase Chain Reaction/methods , Vaginal Smears , Adolescent , Adult , Aged , Alphapapillomavirus/genetics , Colposcopy , Consensus Sequence , Female , Genome, Viral , Humans , Middle Aged , Nucleic Acid Amplification Techniques/methods , Oncogene Proteins, Viral/genetics , Papillomavirus Infections/pathology , Papillomavirus Infections/virology , Reproducibility of Results , Sensitivity and Specificity , Uterine Cervical Dysplasia/pathology , Uterine Cervical Dysplasia/virology , Uterine Cervical Neoplasms/pathology , Uterine Cervical Neoplasms/virology , Uterine Cervicitis/pathology , Uterine Cervicitis/virology
12.
Salus ; 11(1): 16-22, abr. 2007. graf, tab
Article in Spanish | LILACS | ID: lil-502623

ABSTRACT

Se evaluó el efecto antiparasitario del pamoato de pirantel/oxantel y metronidazol y su relación con parámetros hematológicos, en 166 niños entre 3-14 años de la escuela “Ascanio José Velásquez”, Cumaná. Se recolectaron heces frescas a las que se les realizó examen coproparasitológico directo, el método de Ritchie, cultivo de heces en agar, Kato-Katz cuantificado y cuantificación de protozoarios. Además, se extrajo muestras de sangre para las determinaciones hematológicas. Se emplearon métodos clásicos para determinaciones hematológicas. Se emplearon métodos clásicos para determinar eficacia y porcentaje de cura de los medicamentos. Durante las semanas 1, 4 y 9 post tratamiento se determinó el nhpgh de helmintos, número quistes protozoarios y parámetros hematológicos. La prevalencia de parasitosis intestinal fue 85,54 por ciento (142/166), con predominio de poliparasitismo (62,56 por ciento; 103/166); A. lumbricoides y T. trichiura resultaron los helmintos más frecuentes; B. hominis y E. nana fueron los protozoarios más reportados. El grupo erario más afectado fue el de 7-10 años, encontrándose diferencias significativas entre edad y grado de infección para A. lumbricoides (X²= 6,67; p<0,05) y B. hominis (X²= 8,28; p<0,05). La eficacia de los medicamentos fue elevada, con excepción de T. trichiura, frente al cual la eficacia del pamoato de pirantel/oxantel fue inferior a lo esperado. Se encontró diferencias significativas entre los valores de hemoglobina para los niños tratados con pamoato de pirantel/oxantel (F= 3,95; p<0,05) y metronidazol (F= 3,67; p<0,05), y entre los valores de hematocrito para ambos medicamentos (F= 4,21; p<0,05) y (F= 4,00; p<0,05), respectivamente


Subject(s)
Humans , Male , Female , Child , Adolescent , Antiparasitic Agents , Hematologic Agents , Intestinal Diseases, Parasitic , Pediatrics , Venezuela
13.
Kasmera ; 33(2): 142-154, jul.-dic. 2005. tab, graf
Article in Spanish | LILACS | ID: lil-436433

ABSTRACT

Se evaluó la eficacia del pamoato de pirantel/oxantel y la reinfección helmíntica, en niños de ambos sexos y entre 3 y 14 años. Se seleccionaron 289 niños, 123 en Agua Blanca, municipio Montes y 166 en Malariología, municipio Sucre. Se recolectaron muestras fecales frescas, procesadas mediante examen coproparasitológico con solución salina y lugol, y los métodos de Ritchie, cultivo de heces en placas de agar, Kato-Katz cuantitativo y cuantificación de protozoarios. Se realizaron cuatro muestreos post tratamiento (semanas 1, 3, 5 y 10). Los resultados obtenidos se expresaron en tablas de prevalencia, además se aplicaron las pruebas de Ji cuadrado (c²) y ANOVA múltiple a un 95 por ciento de confiabilidad, y se utilizaron métodos clásicos para determinar la eficacia del medicamento, de acuerdo con la disminución del número de hpgh; su efectividad, determinando el porcentaje de casos curados que habían estado infectados al inicio del estudio y la reinfección, calculado como la proporción entre las nuevas infecciones y los casos curados, para cada una de las especies evaluadas. La prevalencia de parasitosis intestinal fue 99,19 por ciento en Agua Blanca y 85,54 por ciento en Malariología. Trichuris trichiura resultó el helminto más prevalente en ambas poblaciones (74,84 por ciento y 50,00 por ciento), y por los protozoarios Blastocystis hominis (73,17 por ciento y 40,96 por ciento). Según el grado de infección helmíntica, hubo predominio de infecciones leves, sin que se observara diferencia significativa por población (F=2,76; p>0,05 para Ascaris lumbricoides, F=2,13; p>0,05 para T. trichiura y F=1,17;p>0,05 para Necator americanus). El grupo de 7 a 10 años fue el más afectado. El pamoato de pirantel/oxantel tuvo mayor eficacia y efectividad frente a A. lumbricoides. Hubo asociación entre las reinfecciones y las comunidades estudiadas (c2=14,82;p>0,05). Las características ambientales de las comunidades podrían favorecer la transmisión helmíntica y propiciar reinf...


Subject(s)
Humans , Male , Female , Child , Anthelmintics/therapeutic use , Intestinal Diseases, Parasitic , Treatment Outcome , Pediatrics , Venezuela
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