ABSTRACT
OBJECTIVE: Ureaplasma urealyticum is one of the organisms most frequently isolated from the amniotic fluid of women with adverse pregnancy. The prevalence of U. urealyticum and U. parvum on samples of amniotic fluid from healthy asymptomatic pregnant women, and whether its detection is associated with P-PROM or preterm birth was investigated. METHODS: Transabdominal amniotic fluid obtained from 121 asymptomatic women at 16-20 weeks of gestation were tested for the detection of Ureaplasma spp., using a selective culture media. A Multiplex-Polymerase Chain Reaction (PCR) method was used for the identification of U. parvum and U. urealyticum. Pregnancy outcomes were obtained after the completion of all testing. RESULTS: Ureaplasma spp. was not identified by culture, but was identified by Multiplex-PCR in four subjects, two corresponding to U. parvum and two to U. urealyticum. The women positive to ureaplasmas had normal labor, and babies born from infected-ureaplasmas pregnant women had normal weight birth. Preterm birth with intact membranes was documented in four women, all negative to ureaplasmas, but associated with gestational hypertension, lost of liquids and low weight birth. CONCLUSIONS: Multiplex-PCR method was more sensitive that culture in detecting ureaplasma organism in amniotic fluid. No association of ureaplasmas with pregnancy outcomes was found.
Subject(s)
Amniotic Fluid/microbiology , Pregnancy Complications, Infectious/etiology , Ureaplasma Infections/complications , Ureaplasma urealyticum/isolation & purification , Female , Humans , Infant, Low Birth Weight , Infant, Newborn , Infant, Premature , Polymerase Chain Reaction , Pregnancy , Pregnancy OutcomeABSTRACT
Se aplicó un método de la reacción en cadena de la polimerasa para la detección temprana de Leptospiras spp. en hemocultivos procedentes de pacientes con sospecha de leptospirosis humana. Los métodos moleculares, y en particular la amplificación del ADN por la reacción en cadena de la polimerasa y sus variantes, constituyen herramientas muy útiles y específicas, utilizadas en la actualidad con esta finalidad. Se lograron identificar por la reacción en cadena de la polimerasa Leptospiras spp. en 41,3 por ciento (33/80) de los hemocultivos a los 7 d de incubación, los que también se clasificaron como positivos por los métodos convencionales. Sin embargo, hubo 20 por ciento (16/80) en los que también por este método se lograron identificar Leptospiras spp., pero por los métodos convencionales resultaron ser negativos. De los hemocultivos 38,7 por ciento (31/80) resultó negativo tanto por la reacción en cadena de la polimerasa como por los métodos convencionales. El uso del método de la reacción en cadena de la polimerasa a partir de hemocultivos, es una alternativa valiosa para la detección temprana y el diagnóstico rápido de Leptospiras spp(AU)
The PCR method for the early detection of Leptospiras spp. in hemocultures from patients suspected of human leptospiros was applied. Molecular methods and, particularly, the amplification of DNA by PCR, and its variants, are very useful and specific tools used nowdays to this end. Leptospiras spp. were identied by means of PCR in 41.3 percent(33/80) of the hemocultures at 7 days of incubation.They were also classified as positive by the conventional methods. However, it was also possible to identify Leptospiras spp. in 20 percent (16/80) by this method, but they proved to be negative by the conventional methods. Of the hemocultures, 38.7 percent (31/80) yielded negative by PCR and by the conventional methods. The use of PCR starting from hemocultures is a valuable alternative for the early detection and rapid diagnosis of Leptospiras spp(AU)
Subject(s)
Leptospira/enzymology , Polymerase Chain Reaction/methodsABSTRACT
Se evaluaron por vez primera en Cuba las tecnologías serológicas rápidas para realizar la pesquisa y confirmación de la leptospirosis humana, que mundialmente están disponibles. Su reconocimiento clínico resulta difícil, el diagnóstico etiológico rápido es de vital importancia Se obtuvieron valores de sensibilidad y especificidad superiores a 90 por ciento en los sistemas confirmatorios y de 85 por ciento en el de pesquisa. De los pacientes graves estudiados por estas tecnologías 50 por ciento fue positivo, además se confirmaron 203 casos pertenecientes a 4 brotes epidémicos, así como 12 enfermos vacunados con vaxSPIRAL®, demostrándose la eficacia de la vacuna (78,1 por ciento). Se desarrolló y aplicó un sistema látex para pesquisa rápida de leptospirosis (LeptoCuba), con excelente sensibilidad, especificidad, reproducibilidad y estabilidad. La aplicación de nuevas tecnologías a la pesquisa y confirmación rápida de la enfermedad, permitió incrementar la positividad y calidad del diagnóstico en el período 2002-2006, fortaleciéndose la vigilancia microbiológica en el país(AU)
For the first time in Cuba the rapid serologic technologies available worldwide were evaluated for the screening and confirmation of human leptospirosis. As its clinical recognition is difficult, the fast etiological diagnosis is of vital importance. Sensitivity and specificity values higher than 90 percent were obtained in the confirmatory systems, and of 85 percent in the screening systems. Of the severe patients studied by these technologies 50 percent were positive. 203 cases corresponding to 4 epidemic outbreaks were confirmed, as well as 12 sick persons vaccinated with vaxSPIRAL®, which demonstrates the vaccine effectiveness (78.1 percent). We developed and applied a latex system for a rapid screening of leptospirosis (LeptoCuba), with an excellent sensitivity, specificity, reproducibility, and stability. The application of new technologies for the screening and fast confirmation of the disease allowed to increase the positivity and quality of the diagnosis from 2000 to 2006, strengthening the microbiological surveillance in the country(AU)
Subject(s)
Humans , Leptospirosis/diagnosis , Serologic TestsABSTRACT
Se aplicó un método de la reacción en cadena de la polimerasa para la detección temprana de Leptospiras spp. en hemocultivos procedentes de pacientes con sospecha de leptospirosis humana. Los métodos moleculares, y en particular la amplificación del ADN por la reacción en cadena de la polimerasa y sus variantes, constituyen herramientas muy útiles y específicas, utilizadas en la actualidad con esta finalidad. Se lograron identificar por la reacción en cadena de la polimerasa Leptospiras spp. en 41,3 por ciento (33/80) de los hemocultivos a los 7 d de incubación, los que también se clasificaron como positivos por los métodos convencionales. Sin embargo, hubo 20 por ciento (16/80) en los que también por este método se lograron identificar Leptospiras spp., pero por los métodos convencionales resultaron ser negativos. De los hemocultivos 38,7 por ciento (31/80) resultó negativo tanto por la reacción en cadena de la polimerasa como por los métodos convencionales. El uso del método de la reacción en cadena de la polimerasa a partir de hemocultivos, es una alternativa valiosa para la detección temprana y el diagnóstico rápido de Leptospiras spp.
The PCR method for the early detection of Leptospiras spp. in hemocultures from patients suspected of human leptospiros was applied. Molecular methods and, particularly, the amplification of DNA by PCR, and its variants, are very useful and specific tools used nowdays to this end. Leptospiras spp. were identied by means of PCR in 41.3 percent(33/80) of the hemocultures at 7 days of incubation.They were also classified as positive by the conventional methods. However, it was also possible to identify Leptospiras spp. in 20 percent (16/80) by this method, but they proved to be negative by the conventional methods. Of the hemocultures, 38.7 percent (31/80) yielded negative by PCR and by the conventional methods. The use of PCR starting from hemocultures is a valuable alternative for the early detection and rapid diagnosis of Leptospiras spp
Subject(s)
Leptospira/enzymology , Polymerase Chain Reaction/methodsABSTRACT
Se evaluaron por vez primera en Cuba las tecnologías serológicas rápidas para realizar la pesquisa y confirmación de la leptospirosis humana, que mundialmente estßn disponibles. Su reconocimiento clínico resulta difícil, el diagnóstico etiológico rápido es de vital importancia Se obtuvieron valores de sensibilidad y especificidad superiores a 90 por ciento en los sistemas confirmatorios y de 85 por ciento en el de pesquisa. De los pacientes graves estudiados por estas tecnologías 50 por ciento fue positivo, además se confirmaron 203 casos pertenecientes a 4 brotes epidémicos, así como 12 enfermos vacunados con vaxSPIRAL®, demostrándose la eficacia de la vacuna (78,1 por ciento). Se desarrolló y aplicó un sistema látex para pesquisa rápida de leptospirosis (LeptoCuba), con excelente sensibilidad, especificidad, reproducibilidad y estabilidad. La aplicación de nuevas tecnologías a la pesquisa y confirmación rápida de la enfermedad, permitió incrementar la positividad y calidad del diagnóstico en el período 2002-2006, fortaleciéndose la vigilancia microbiológica en el país.
For the first time in Cuba the rapid serologic technologies available worldwide were evaluated for the screening and confirmation of human leptospirosis. As its clinical recognition is difficult, the fast etiological diagnosis is of vital importance. Sensitivity and specificity values higher than 90 percent were obtained in the confirmatory systems, and of 85 percent in the screening systems. Of the severe patients studied by these technologies 50 percent were positive. 203 cases corresponding to 4 epidemic outbreaks were confirmed, as well as 12 sick persons vaccinated with vaxSPIRAL®, which demonstrates the vaccine effectiveness (78.1 percent). We developed and applied a latex system for a rapid screening of leptospirosis (LeptoCuba), with an excellent sensitivity, specificity, reproducibility, and stability. The application of new technologies for the screening and fast confirmation of the disease allowed to increase the positivity and quality of the diagnosis from 2000 to 2006, strengthening the microbiological surveillance in the country
Subject(s)
Humans , Leptospirosis/diagnosis , Serologic TestsABSTRACT
For the first time in Cuba the rapid serologic technologies available worldwide were evaluated for the screening and confirmation of human leptospirosis. As its clinical recognition is difficult, the fast etiological diagnosis is of vital importance. Sensitivity and specificity values higher than 90 % were obtained in the confirmatory systems, and of 85 % in the screening systems. Of the severe patients studied by these technologies 50 % were positive. 203 cases corresponding to 4 epidemic outbreaks were confirmed, as well as 12 sick persons vaccinated with vaxSPIRAL, which demonstrates the vaccine effectiveness (78.1%). We developed and applied a latex system for a rapid screening of leptospirosis (LeptoCuba), with an excellent sensitivity, specificity, reproducibility, and stability. The application of new technologies for the screening and fast confirmation of the disease allowed to increase the positivity and quality of the diagnosis from 2000 to 2006, strengthening the microbiological surveillance in the country.
Subject(s)
Leptospirosis/diagnosis , Reagent Kits, Diagnostic , Serologic Tests/trends , Animal Diseases/diagnosis , Animals , Antibodies, Bacterial/blood , Bacterial Vaccines , Clinical Trials as Topic , Cuba/epidemiology , Disease Outbreaks , Hemagglutination Tests , Humans , Immunoglobulin M/blood , Latex Fixation Tests , Leptospira/classification , Leptospira/immunology , Leptospirosis/epidemiology , Leptospirosis/prevention & control , Leptospirosis/veterinary , Reproducibility of Results , Rural Population , Sensitivity and Specificity , Seroepidemiologic Studies , Serotyping , Time Factors , Urban Population , VaccinationABSTRACT
The PCR method for the early detection of Leplospiras spp. in hemocultures from patients suspected of human leptospiros was applied. Molecular methods and, particularly, the amplification of DNA by PCR, and its variants, are very useful and specific tools used nowdays to this end. Leptospiras spp. were identied by means of PCR in 41.3 % (33/80) of the hemocultures at 7 days of incubation. They were also classified as positive by the conventional methods. However, it was also possible to identify Leptospiras spp. in 20% (16/80) by this method, but they proved to be negative by the conventional methods. Of the hemocultures, 38.7% (31/80) yielded negative by PCR and by the conventional methods. The use of PCR starting from hemocultures is a valuable alternative for the early detection and rapid diagnosis of Leptospiras spp.
Subject(s)
Bacteremia/diagnosis , Leptospira/isolation & purification , Leptospirosis/diagnosis , Polymerase Chain Reaction/methods , Bacteremia/microbiology , Early Diagnosis , False Negative Reactions , Humans , Leptospira/genetics , Leptospirosis/blood , Leptospirosis/microbiology , Reproducibility of ResultsABSTRACT
Se aplicó un sistema ELISA cuantitativo para la detección de anticuerpos IgG en respuesta a la vacuna cubana contra la leptospirosis humana (vax-SPIRAL), a sueros de 930 voluntarios, 483 inmunizados con vax-SPIRAL y 447 con una vacuna contra la hepatitis B. Fueron estudiadas las muestras tomadas antes de comenzar el estudio, a los 21 d de aplicada la segunda dosis y al cabo del año. Se pudo observar una alta seroprevalencia de anticuerpos a los serovares vacunales antes de comenzar el estudio, un incremento al doble del valor inicial de la respuesta a los 21 d de la segunda dosis de la vacuna antileptospirósica en 45 por ciento de los individuos, existiendo diferencias marcadamente significativas (p= 0,000000) entre ambos grupos. Se detectó respuesta a los 3 serovares leptospirales de forma similar. Al cabo del año los niveles de anticuerpos caen, sin embargo, hay 23,1 por ciento de individuos que logran duplicar su nivel de anticuerpos respecto al momento inicial, manteniéndose la diferencia significativa con el grupo que recibió la vacuna contra hepatitis B. Se recomendó la continuación de la aplicación de este inmunógeno cubano a los principales grupos de riesgo
Subject(s)
Humans , Adult , Antibodies, Anti-Idiotypic , Immunoglobulin G , Leptospira , Leptospirosis , Vaccines , Enzyme-Linked Immunosorbent AssayABSTRACT
Se aplicó un sistema ELISA cuantitativo para la detección de anticuerpos IgG en respuesta a la vacuna cubana contra la leptospirosis humana (vax-SPIRAL), a sueros de 930 voluntarios, 483 inmunizados con vax-SPIRAL y 447 con una vacuna contra la hepatitis B. Fueron estudiadas las muestras tomadas antes de comenzar el estudio, a los 21 d de aplicada la segunda dosis y al cabo del año. Se pudo observar una alta seroprevalencia de anticuerpos a los serovares vacunales antes de comenzar el estudio, un incremento al doble del valor inicial de la respuesta a los 21 d de la segunda dosis de la vacuna antileptospirósica en 45 por ciento de los individuos, existiendo diferencias marcadamente significativas (p= 0,000000) entre ambos grupos. Se detectó respuesta a los 3 serovares leptospirales de forma similar. Al cabo del año los niveles de anticuerpos caen, sin embargo, hay 23,1 por ciento de individuos que logran duplicar su nivel de anticuerpos respecto al momento inicial, manteniéndose la diferencia significativa con el grupo que recibió la vacuna contra hepatitis B. Se recomendó la continuación de la aplicación de este inmunógeno cubano a los principales grupos de riesgo(AU)
Subject(s)
Humans , Adult , Leptospira/immunology , Leptospirosis/immunology , Vaccines/immunology , Antibodies, Anti-Idiotypic , Immunoglobulin G , Enzyme-Linked Immunosorbent Assay/methodsABSTRACT
A quantitative ELISA for the detection of IgG antibodies in response to the Cuban vaccine against the human leptospirosis (vax-SPIRAL) was applied to 930 volunteers'sera, 483 immunized with vax-SPIRAL and 447 with a vaccine against hepatitis B. Samples were taken before beginning the study, 21 days after the second dose and a year later. A high seroprevalence of antibodies to the vaccine serovars was observed before beginning the study. The initial value of the response doubled at 21 days of the second dose in 45 % of the individuals. There were markedly significant differences (p=0,000000) between both groups. A similar response to the three leptospiral serovars was found. After the year the levels of antibodies decreased; however, 23.1% of the individuals were able to duplicate their level of antibodies regarding the initial moment, and the significant difference with the group that received the vaccine against hepatitis B still existed. The application of this Cuban immunogen amomg the main risk groups was recommended.
