RESUMEN
BACKGROUND: Preterm prelabor rupture of the membranes (PPROM) is frequently complicated by intraamniotic inflammatory processes such as intraamniotic infection and sterile intraamniotic inflammation. Antibiotic therapy is recommended to patients with PPROM to prolong the interval between this complication and delivery (latency period), reduce the risk of clinical chorioamnionitis, and improve neonatal outcome. However, there is a lack of information regarding whether the administration of antibiotics can reduce the intensity of the intraamniotic inflammatory response or eradicate microorganisms in patients with PPROM. OBJECTIVE: The first aim of the study was to determine whether antimicrobial agents can reduce the magnitude of the intraamniotic inflammatory response in patients with PPROM by assessing the concentrations of interleukin-6 in amniotic fluid before and after antibiotic treatment. The second aim was to determine whether treatment with intravenous clarithromycin changes the microbial load of Ureaplasma spp DNA in amniotic fluid. STUDY DESIGN: A retrospective cohort study included patients who had (1) a singleton gestation, (2) PPROM between 24+0 and 33+6 weeks, (3) a transabdominal amniocentesis at the time of admission, and (4) intravenous antibiotic treatment (clarithromycin for patients with intraamniotic inflammation and benzylpenicillin/clindamycin in the cases of allergy in patients without intraamniotic inflammation) for 7 days. Follow-up amniocenteses (7th day after admission) were performed in the subset of patients with a latency period lasting longer than 7 days. Concentrations of interleukin-6 were measured in the samples of amniotic fluid with a bedside test, and the presence of microbial invasion of the amniotic cavity was assessed with culture and molecular microbiological methods. Intraamniotic inflammation was defined as a bedside interleukin-6 concentration ≥745 pg/mL in the samples of amniotic fluid. Intraamniotic infection was defined as the presence of both microbial invasion of the amniotic cavity and intraamniotic inflammation; sterile intraamniotic inflammation was defined as the presence of intraamniotic inflammation without microbial invasion of the amniotic cavity. RESULTS: A total of 270 patients with PPROM were included in this study: 207 patients delivered within 7 days and 63 patients delivered after 7 days of admission. Of the 63 patients who delivered after 7 days following the initial amniocentesis, 40 underwent a follow-up amniocentesis. Patients with intraamniotic infection (n = 7) and sterile intraamniotic inflammation (n = 7) were treated with intravenous clarithromycin. Patients without either microbial invasion of the amniotic cavity or intraamniotic inflammation (n = 26) were treated with benzylpenicillin or clindamycin. Treatment with clarithromycin decreased the interleukin-6 concentration in amniotic fluid at the follow-up amniocentesis compared to the initial amniocentesis in patients with intraamniotic infection (follow-up: median, 295 pg/mL, interquartile range [IQR], 72-673 vs initial: median, 2973 pg/mL, IQR, 1750-6296; P = .02) and in those with sterile intraamniotic inflammation (follow-up: median, 221 pg/mL, IQR 118-366 pg/mL vs initial: median, 1446 pg/mL, IQR, 1300-2941; P = .02). Samples of amniotic fluid with Ureaplasma spp DNA had a lower microbial load at the time of follow-up amniocentesis compared to the initial amniocentesis (follow-up: median, 1.8 × 104 copies DNA/mL, 2.9 × 104 to 6.7 × 108 vs initial: median, 4.7 × 107 copies DNA/mL, interquartile range, 2.9 × 103 to 3.6 × 107; P = .03). CONCLUSION: Intravenous therapy with clarithromycin was associated with a reduction in the intensity of the intraamniotic inflammatory response in patients with PPROM with either intraamniotic infection or sterile intraamniotic inflammation. Moreover, treatment with clarithromycin was related to a reduction in the load of Ureaplasma spp DNA in the amniotic fluid of patients with PPROM <34 weeks of gestation.
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Antibacterianos/uso terapéutico , Infecciones Bacterianas/prevención & control , Corioamnionitis/prevención & control , Claritromicina/uso terapéutico , Clindamicina/uso terapéutico , Rotura Prematura de Membranas Fetales , Penicilina G/uso terapéutico , Adulto , Líquido Amniótico/química , Infecciones Bacterianas/etiología , Corioamnionitis/etiología , Estudios de Cohortes , ADN Bacteriano/análisis , Femenino , Humanos , Interleucina-6/análisis , Embarazo , Estudios Retrospectivos , Ureaplasma/genéticaRESUMEN
BACKGROUND: To determine the association between microbial invasion of the amniotic cavity (MIAC) and the presence of Lactobacillus crispatus- or Lactobacillus iners-dominated cervical microbiota in pregnancies with preterm prelabor rupture of membrane. Next, to assess the relationship between the presence of L. crispatus- or L. iners-dominated cervical microbiota and short-term neonatal morbidity. METHOD: A total of 311 women were included. Cervical samples were obtained using a Dacron polyester swab and amniotic fluid samples were obtained by transabdominal amniocentesis. Bacterial DNA, L. crispatus, and L. iners in the cervical samples were assessed by PCR. Cervical microbiota was assigned as L. crispatus- or L. iners-dominated when the relative abundance of L. crispatus or L. iners was ≥50% of the whole cervical microbiota, respectively. RESULTS: Women with MIAC showed a lower rate of L. crispatus-dominated cervical microbiota (21% vs. 39%; p = 0.003) than those without MIAC. Lactobacillus crispatus-dominated cervical microbiota was associated with a lower rate of early-onset sepsis (0% vs. 5%; p = 0.02). CONCLUSIONS: The presence of L. crispatus-dominated cervical microbiota in women with preterm prelabor rupture of membrane was associated with a lower risk of intra-amniotic complications and subsequent development of early-onset sepsis of newborns.
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Amniocentesis/métodos , Líquido Amniótico/microbiología , Corioamnionitis/microbiología , Rotura Prematura de Membranas Fetales/microbiología , Lactobacillus crispatus , Lactobacillus , Cuello del Útero/microbiología , Chlamydia trachomatis , Femenino , Humanos , Recién Nacido , Microbiota , Mycoplasma hominis , Trabajo de Parto Prematuro , Embarazo , Estudios Retrospectivos , UreaplasmaRESUMEN
The aim of this study was to test the detection performance of the cpsA, lytA and ply genes through qPCR in the identification of Streptococcus pneumoniae in respiratory tract samples. Specificity was tested on a panel of 128 streptococci and other bacteria DNA samples. The qPCR assay was tested on a total of 51 respiratory tract samples from patients with community-acquired pneumonia (CAP). The specificity of the cpsA, lytA and ply genes was 100%, 100%, and 86%, respectively. The quantitative assessment, based on lytA, determined a cutoff value of ~2x104, 4x102 and 4x102 DNA copies per 1 mL of valid sputum, tracheal aspirate and bronchial aspirate samples, respectively. The results from the present study suggest that qPCR detection of all three genes would be optimal in the accurate detection of Streptococcus pneumoniae.
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Infecciones Comunitarias Adquiridas , Neumonía Neumocócica , Reacción en Cadena en Tiempo Real de la Polimerasa , Infecciones Comunitarias Adquiridas/diagnóstico , Infecciones Comunitarias Adquiridas/microbiología , ADN Bacteriano/genética , Humanos , Neumonía Neumocócica/diagnóstico , Neumonía Neumocócica/microbiología , Sistema Respiratorio/microbiología , Sensibilidad y Especificidad , Streptococcus pneumoniae/genéticaRESUMEN
PROBLEM: To determine the changes of pentraxin 3 (PTX3) level in noninvasively obtained cervical fluid samples from women with preterm prelabor rupture of membranes (PPROM) based on the presence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI), and intra-amniotic infection (the presence of both MIAC and IAI). METHODS OF STUDY: A total of 160 women with PPROM were included. Cervical fluid samples were obtained using a Dacron polyester swab and amniotic fluid samples were obtained by transabdominal amniocentesis. Cervical fluid PTX3 levels were assessed using enzyme-linked immunosorbent assay. RESULTS: PTX3 was found in all the cervical fluid samples and its levels were higher in women with MIAC, IAI, and intra-amniotic infection than in women without these conditions. When the women were categorized into four subgroups based on the presence of MIAC and/or IAI, women with intra-amniotic infection had higher cervical fluid PTX3 levels than those with sterile IAI (IAI alone), colonization (MIAC alone), or no MIAC or IAI. A cervical fluid PTX3 level of 11 ng/mL was the best value for identifying the presence of intra-amniotic infection in women with PPROM. CONCLUSIONS: PTX3 is a constituent of cervical fluid of women with PPROM. Cervical fluid PTX3 level reflects the situation in the intra-amniotic compartments of women with PPROM. Cervical fluid PTX3 is a potential marker for the noninvasive identification of intra-amniotic infection in PPROM.
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Proteína C-Reactiva/metabolismo , Cuello del Útero/metabolismo , Corioamnionitis/metabolismo , Rotura Prematura de Membranas Fetales/metabolismo , Componente Amiloide P Sérico/metabolismo , Líquido Amniótico/microbiología , Biomarcadores/metabolismo , Corioamnionitis/diagnóstico , Corioamnionitis/microbiología , Ensayo de Inmunoadsorción Enzimática , Femenino , Rotura Prematura de Membranas Fetales/diagnóstico , Rotura Prematura de Membranas Fetales/microbiología , Humanos , Valor Predictivo de las Pruebas , Embarazo , Estudios Retrospectivos , Regulación hacia ArribaRESUMEN
Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (â¼30, â¼45, and â¼80 kb). Sequencing revealed three types of plasmids (A to C) with the Tn4401a transposon. Type A plasmids comprised an IncR backbone and a KPC-2-encoding multidrug resistance (MDR) region. Type B plasmids were derivatives of type A plasmids carrying an IncN3-like segment, while type C plasmids were IncP6 plasmids sharing the same KPC-2-encoding MDR region with type A and B plasmids.
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Citrobacter freundii/genética , Escherichia coli/genética , Klebsiella pneumoniae/genética , Morganella morganii/genética , Plásmidos/química , beta-Lactamasas/genética , Antibacterianos/uso terapéutico , Secuencia de Bases , Carbapenémicos/uso terapéutico , Citrobacter freundii/efectos de los fármacos , Citrobacter freundii/enzimología , Citrobacter freundii/aislamiento & purificación , República Checa/epidemiología , Elementos Transponibles de ADN , Farmacorresistencia Bacteriana Múltiple/genética , Infecciones por Enterobacteriaceae/tratamiento farmacológico , Infecciones por Enterobacteriaceae/epidemiología , Infecciones por Enterobacteriaceae/microbiología , Escherichia coli/efectos de los fármacos , Escherichia coli/enzimología , Escherichia coli/aislamiento & purificación , Expresión Génica , Hospitales , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Klebsiella pneumoniae/efectos de los fármacos , Klebsiella pneumoniae/enzimología , Klebsiella pneumoniae/aislamiento & purificación , Pruebas de Sensibilidad Microbiana , Morganella morganii/efectos de los fármacos , Morganella morganii/enzimología , Morganella morganii/aislamiento & purificación , Sistemas de Lectura Abierta , Plásmidos/clasificación , Plásmidos/metabolismo , Análisis de Secuencia de ADN , beta-Lactamasas/metabolismoRESUMEN
OBJECTIVE: To determine changes in the intraamniotic environment during the latency period using paired amniotic and gastric fluid samples in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). METHODS: A total of 34 women with singleton pregnancies complicated by PPROM prior to 34 weeks were included in the study. Amniotic fluid was obtained by transabdominal amniocentesis at the time of admission. Immediately after delivery, umbilical cord blood and gastric fluid were obtained. RESULT: Microorganisms in amniotic and gastric fluid samples were found in 38% and 59% of women, respectively. Bedside IL-6 levels were higher in amniotic than in gastric fluid in pregnancies without fetal inflammatory response syndrome (FIRS) (263 pg/mL vs. 50 pg/mL; p < 0.0001), but not in pregnancies with FIRS (318 pg/mL vs. 444 pg/mL; p = 0.91). Funisitis and FIRS was associated with the highest bedside IL-6 levels in gastric fluid. A gastric fluid bedside IL-6 level of 275 pg/mL was found to be the ideal cutoff value to predict funisitis and FIRS. CONCLUSIONS: The microbial and inflammatory status of the intraamniotic compartment changes during the latency period in PPROM. Bedside IL-6 assessment of gastric fluid may be useful in the rapid diagnosis of funisitis and FIRS.
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Líquido Amniótico/química , Sangre Fetal/química , Rotura Prematura de Membranas Fetales , Jugo Gástrico/química , Adulto , Amniocentesis , Líquido Amniótico/microbiología , Biomarcadores/análisis , Líquidos Corporales , Chlamydia trachomatis , Corioamnionitis , Femenino , Jugo Gástrico/microbiología , Humanos , Recién Nacido , Inflamación , Interleucina-6/análisis , Mycoplasma hominis , Embarazo , Estudios Prospectivos , Estómago/microbiología , Síndrome , UreaplasmaRESUMEN
BackgroundTo characterize the influence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI) on the intensity of the fetal inflammatory response and the association between the presence of the fetal inflammatory response syndrome (FIRS) and short-term neonatal morbidity in the preterm prelabor rupture of membranes (PPROM) between the gestational ages of 34 and 37 weeks.MethodsOne hundred and fifty-nine women were included in the study. The umbilical cord blood interleukin (IL)-6 concentrations were determined using enzyme-linked immunosorbent assay kits. FIRS was defined based on the umbilical cord blood IL-6 concentration and the presence of funisitis and/or chorionic plate vasculitis.ResultsWomen with both MIAC and IAI had the highest median umbilical cord blood IL-6 concentrations and highest rates of FIRS. Women with FIRS had the higher rates of early-onset sepsis and intraventricular hemorrhage grades I and II when FIRS was characterized based on the umbilical cord blood IL-6 concentrations and the histopathological findings.ConclusionThe presence of both MIAC and IAI was associated with a higher fetal inflammatory response and a higher rate of FIRS. Different aspects of short-term neonatal morbidity were related to FIRS when defined by umbilical cord blood IL-6 concentrations and the histopathology of the placenta.
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Líquido Amniótico/microbiología , Corioamnionitis/microbiología , Rotura Prematura de Membranas Fetales/microbiología , Inflamación/microbiología , Interleucina-6/sangre , Adulto , Chlamydia trachomatis , Ensayo de Inmunoadsorción Enzimática , Femenino , Sangre Fetal/química , Edad Gestacional , Humanos , Recién Nacido , Mycoplasma hominis , Placenta/patología , Embarazo , Resultado del Embarazo , Estudios Prospectivos , Ureaplasma/metabolismo , Vasculitis/microbiologíaRESUMEN
INTRODUCTION: We evaluated the levels of cell-free nuclear DNA (nDNA) and cell-free mitochondrial DNA (mtDNA) in the amniotic fluid supernatant from pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on evidence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI). MATERIAL AND METHODS: A total of 155 women with PPROM were included in this study. Amniotic fluid samples were obtained by transabdominal amniocentesis. The levels of cell-free nDNA and mtDNA in the amniotic fluid supernatant were assessed and quantified by real-time polymerase chain reaction. RESULTS: The levels of cell-free nDNA and mtDNA were higher in women with MIAC and IAI than in women without these conditions (nDNA: with MIAC: median 3.9 × 104 genome equivalent [GE]/mL vs without MIAC: median 1.2 × 104 GE/mL, with IAI: median: 5.3 × 104 GE/mL vs without IAI: median 1.2 × 104 GE/mL; mtDNA: with MIAC: median 9.2 × 105 GE/mL vs without MIAC: median 2.5 × 105 GE/mL, with IAI: median 1.1 × 106 GE/mL vs without IAI: median 2.5 × 105 ; all P values ≤ 0.01). Women with the microbial-associated IAI showed the highest levels of cell-free nDNA and mtDNA. CONCLUSIONS: Cell-free nDNA and mtDNA are constituents of the amniotic fluid supernatant from PPROM pregnancies. Both cell-free nDNA and mtDNA are involved in the intra-amniotic inflammatory response in women with PPROM.
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Líquido Amniótico/metabolismo , Infecciones Bacterianas/metabolismo , Ácidos Nucleicos Libres de Células/metabolismo , Corioamnionitis/metabolismo , ADN Mitocondrial/metabolismo , Rotura Prematura de Membranas Fetales/metabolismo , Inflamación/metabolismo , Adulto , Amniocentesis , Líquido Amniótico/microbiología , Chlamydia trachomatis , Estudios de Cohortes , Técnicas de Cultivo , Femenino , Edad Gestacional , Humanos , Interleucina-6/metabolismo , Mycoplasma hominis , Reacción en Cadena de la Polimerasa , Embarazo , ARN Ribosómico 16S/análisis , Reacción en Cadena en Tiempo Real de la Polimerasa , Estudios Retrospectivos , UreaplasmaRESUMEN
Human polyomaviruses such as JC polyomavirus and BK polyomavirus have long been well known pathogens of immunocompromised patients. Several new members of this viral family have been described during the last decade. Human polyomavirus 9 seems to be a novel pathogen of transplanted patients according to some studies. The aim of our study was to determine the presence of human polyomavirus 9 in patients after kidney or stem cell transplantation (SCT) at the University Hospital in Hradec Kralove, Czech Republic. Overall 100 patients, 65 after kidney transplantation and 35 after SCT, were included into the study. At least three follow-up samples from each patient were examined for human polyomavirus 9 DNA presentation with the two previously described in-house PCR protocols. Despite the frequent reactivation of human CMV (14.3% in kidney transplantation and 63.3% after SCT) or BK polyomavirus in our patient group, there was no positivity for human polyomavirus 9 either in blood samples or urine samples. One of the possible reasons for this discrepancy versus previous published studies could be a relatively low proportion of patients treated by induction therapy before kidney transplantation in our study cohort.
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Huésped Inmunocomprometido , Polyomaviridae/genética , Polyomaviridae/aislamiento & purificación , Infecciones por Polyomavirus/epidemiología , Infecciones por Polyomavirus/virología , Adulto , Anciano , Estudios de Cohortes , República Checa/epidemiología , ADN Viral/genética , Femenino , Hospitales Universitarios , Humanos , Trasplante de Riñón/efectos adversos , Masculino , Persona de Mediana Edad , Reacción en Cadena de la Polimerasa , Polyomaviridae/patogenicidad , Trasplante de Células Madre/efectos adversos , Adulto JovenRESUMEN
Polyomaviruses belong to a group of viruses that has recently attracted the attention of many research groups. During 35 years, JC and BK viruses, known pathogens in immunocompromised patients, seemed to be the only human polyomaviruses. But in 2007, two other polyomaviruses, WU and KI, were isolated whose pathogenicity is still a matter of discussion. A year later, another human polyomavirus, associated with Merkel cell carcinoma, was identified, and seven more were described by the end of 2014. Some of them were found to be related to various diseases, others seem to be a part of the normal skin and mucosal microbiome. The article summarizes basic information about all so far described human polyomaviruses.
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Infecciones por Polyomavirus , Poliomavirus , HumanosRESUMEN
OBJECTIVES: The aim was to introduce a diagnostic method for detecting variants of hepatitis C virus (HCV) with protease NS3 resistance primarily to simeprevir (Q80K mutation in HCV genotype 1a) and its subsequent use in routine practice. MATERIAL AND METHODS: The detection of HCV resistance-associated variants in the NS3 protease gene by sequence analysis was introduced in the molecular biology laboratory of University Hospital Hradec Kralove in 2015. The primers were designed by sequence analysis software Custom Primers - OligoPerfect™ Designer. The method was optimized for HCV genotype 1a. The search for variants was performed using two programs. RESULTS: A total of 16 patients with genotype 1a chronic hepatitis C have been examined since 2015. In five of them, the Q80K variant was detected. CONCLUSION: The development of resistance to antiviral therapy for chronic hepatitis C gained importance after the introduction of direct-acting antivirals. Given the relatively high prevalence of the Q80K mutation in HCV genotype 1a, it is crucial to confirm its presence or absence before the therapy is initiated. The reported method enables clear and early detection of the Q80K mutation.
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Farmacorresistencia Viral/genética , Hepacivirus/genética , Hepatitis C/diagnóstico , Hepatitis C/virología , Proteínas no Estructurales Virales/genética , Virología/métodos , Humanos , Mutación/genéticaRESUMEN
Common variable immunodeficiency (CVID) is the most frequent clinically relevant primary immunodeficiency and shows enormous heterogeneity in clinical presentation. Despite clinical immunodeficiency, opportunistic infections are not a typical manifestation of CVID. A retrospective study of 32 patients followed up for 335 patient-years was performed to determine the frequency of cytomegalovirus (CMV) disease. Symptomatic CMV infection was documented in 3 CVID patients. Patients No. 1 and 2 suffered from CMV pneumonia, with complications due to atypical mycobacteriosis in patient No. 1. Patient No. 3 suffered from CMV enteritis. A history of cancer and chronic hepatitis C infection (patient No. 1), immunosuppressive therapy for interstitial lung disease (patient No. 2) and serious enteropathy complicated with malnutrition (patient No. 3) may have contributed to the complications despite only mild abnormalities in T-cell subpopulations. The direct detection of CMV in bronchoalveolar lavage, stool or tissue samples was the most beneficial diagnostic laboratory method, whereas the detection of CMV DNA in blood did not produce positive results. Adequate treatment of CMV disease led to significant clinical improvement in all 3 patients. The frequency of CMV disease appears to be higher than previously described. In our experience, the probability of opportunistic infections in CVID patients increases with secondary comorbidities and their management.
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Inmunodeficiencia Variable Común/diagnóstico , Infecciones por Citomegalovirus/diagnóstico , ADN Viral/aislamiento & purificación , Enteritis/diagnóstico , Infecciones por Mycobacterium no Tuberculosas/diagnóstico , Neumonía Viral/diagnóstico , Líquido del Lavado Bronquioalveolar/virología , Inmunodeficiencia Variable Común/complicaciones , Inmunodeficiencia Variable Común/patología , Inmunodeficiencia Variable Común/virología , Citomegalovirus , Infecciones por Citomegalovirus/complicaciones , Infecciones por Citomegalovirus/patología , Infecciones por Citomegalovirus/virología , Enteritis/complicaciones , Enteritis/patología , Enteritis/virología , Heces/virología , Femenino , Humanos , Persona de Mediana Edad , Infecciones por Mycobacterium no Tuberculosas/complicaciones , Infecciones por Mycobacterium no Tuberculosas/patología , Infecciones por Mycobacterium no Tuberculosas/virología , Neumonía Viral/complicaciones , Neumonía Viral/patología , Neumonía Viral/virologíaRESUMEN
OBJECTIVE: The objective of the study was to determine the diagnostic indices and predictive values by bedside assessment of amniotic fluid interleukin-6 (IL-6) concentration in the identification of microbial invasion of the amniotic cavity (MIAC) and/or histological chorioamnionitis (HCA) in patients with preterm prelabor rupture of membranes. STUDY DESIGN: One hundred twenty-four women with singleton pregnancies were included in this study. The amniotic fluid was sampled by transabdominal amniocentesis at the time of admission. IL-6 concentrations were assessed with an immunoassay. RESULTS: The presence of MIAC, HCA, or the coexistence of both was associated with higher amniotic fluid concentrations of IL-6 in both a crude and adjusted analysis. The amniotic fluid concentration of IL-6 of 1000 pg/mL was determined to be the best cutoff value for the prediction of MIAC (sensitivity of 50%, specificity of 95%, positive predictive value of 82%, negative predictive value of 81%, and likelihood ratio of 8.4) or both MIAC and HCA (sensitivity of 60%, specificity of 94%, positive predictive value of 75%, negative predictive value of 88%, and likelihood ratio of 9.4). CONCLUSION: The bedside assessment of amniotic fluid IL-6 seems to be an easy, rapid, and inexpensive method for the prediction of MIAC or both MIAC and HCA in pregnancies complicated by preterm prelabor rupture of membranes.
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Amniocentesis , Líquido Amniótico/metabolismo , Corioamnionitis/diagnóstico , Interleucina-6/metabolismo , Infecciones por Mycoplasma/diagnóstico , Sistemas de Atención de Punto , Complicaciones Infecciosas del Embarazo/diagnóstico , Adolescente , Adulto , Líquido Amniótico/microbiología , Biomarcadores/metabolismo , Infecciones por Chlamydia/diagnóstico , Chlamydia trachomatis/aislamiento & purificación , Corioamnionitis/metabolismo , Femenino , Rotura Prematura de Membranas Fetales/metabolismo , Humanos , Mycoplasma hominis/aislamiento & purificación , Valor Predictivo de las Pruebas , Embarazo , Complicaciones Infecciosas del Embarazo/metabolismo , Estudios Prospectivos , Sensibilidad y Especificidad , Infecciones por Ureaplasma/diagnóstico , Infecciones por Ureaplasma/metabolismo , Adulto JovenRESUMEN
OBJECTIVE: We sought to determine the influence of microbial invasion of the amniotic cavity (MIAC) and acute histologic chorioamnionitis (HCA) on the intensity of the intraamniotic inflammatory response and neonatal morbidity in preterm prelabor rupture of membranes (PPROM) between 34-37 weeks. STUDY DESIGN: This study included 99 women with singleton pregnancies complicated by PPROM between the gestational ages of 34-37 weeks. Amniocenteses were performed at the time of admission, and MIAC and amniotic fluid interleukin-6 concentrations were determined. After delivery, the placenta was evaluated for the presence of HCA. RESULTS: Women with both MIAC and HCA had the highest intraamniotic inflammatory response, which was mediated by interleukin-6 concentrations (both MIAC and HCA: median 2164.0 pg/mL; HCA alone: median 654.8 pg/mL; MIAC alone: median 784.1 pg/mL; neither MIAC nor HCA: median 383.0 pg/mL; P < .0001) and the highest incidence of newborns with early-onset sepsis (P = .02). CONCLUSION: Both MIAC and HCA affect the intensity of the intraamniotic inflammatory response and the incidence of early-onset sepsis following PPROM between 34-37 weeks. The intensity of the intraamniotic inflammatory response should be considered in the clinical management of PPROM between 34-37 weeks.
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Líquido Amniótico/química , Líquido Amniótico/microbiología , Corioamnionitis/patología , Rotura Prematura de Membranas Fetales , Interleucina-6/análisis , Sepsis/epidemiología , Adulto , Amniocentesis , Femenino , Edad Gestacional , Humanos , Recién Nacido , Enfermedades del Recién Nacido/diagnóstico , Enfermedades del Recién Nacido/epidemiología , Valor Predictivo de las Pruebas , Embarazo , Estudios Prospectivos , Sensibilidad y Especificidad , Sepsis/diagnóstico , Adulto JovenRESUMEN
BACKGROUND: The aims of this work were to replace the obsolete PCR method for the laboratory diagnosis of the acute form of leptospirosis using the G1, G2 and B64 I, B64 II primers, and to improve the PCR detection time. METHODS: We introduced a real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 of pathogenic Leptospira into our laboratory diagnosis of the acute form of leptospirosis. The positive and negative analytical specificities of the real-time PCR method were both equal to 100%; the detection limit was determined to be 1-5 genome copies/1 ml of liquid biological material. The method was further validated on 230 laboratory strains of leptospires. RESULTS: All laboratory strains of pathogenic Leptospira were evaluated as LipL32-positive and all non-pathogenic strains as LipL32-negative. In addition, 455 biological materials (253 plasma, 121 urine, 72 cerebrospinal fluid (CSF), 7 bronchoalveolar lavage, and 2 sputum) from 295 patients with suspected leptospirosis were examined. From this set of patients, 9 were evaluated to be LipL32-positive, from 15 positive biological materials (10 urine, 4 blood plasma, and 1 CSF). CONCLUSIONS: This real-time PCR method for the detection of the gene encoding the surface lipoprotein LipL32 is a reliable, sensitive, and rapid method for the detection of the acute form of leptospirosis.
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Proteínas de la Membrana Bacteriana Externa/genética , Técnicas Bacteriológicas/métodos , Leptospira/aislamiento & purificación , Leptospirosis/diagnóstico , Lipoproteínas/genética , Técnicas de Diagnóstico Molecular/métodos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Adulto , Anciano , Técnicas de Laboratorio Clínico/métodos , Femenino , Humanos , Leptospira/genética , Leptospirosis/microbiología , Masculino , Persona de Mediana Edad , Sensibilidad y Especificidad , Factores de Tiempo , Adulto JovenRESUMEN
OBJECTIVE: The objective of the study was to determine whether the bacterial load of genital mycoplasmas and gestation age are related to intraamniotic inflammatory response using the amniotic fluid levels of 18 inflammatory mediators. STUDY DESIGN: A prospective study of 145 women with singleton pregnancies complicated by preterm prelabor rupture of membranes between 24(0/7) and 36(6/7) weeks was conducted. Amniotic fluid was obtained from all women by transabdominal amniocentesis. The amounts of genital mycoplasma deoxyribonucleic acid were determined using the threshold cycle value and relative and absolute quantification techniques. A panel of multiple proteins was analyzed simultaneously using multiplex technology. RESULTS: Twenty-four women with the presence of genital mycoplasmas in amniotic fluid were included in the final analyses. The concentrations of 9 of the 18 evaluated proteins in the amniotic fluid correlated with bacterial load of genital mycoplasmas independent of the quantification technique used. CONCLUSION: The intensity of intraamniotic inflammatory response to genital mycoplasmas decreased with gestational age.
Asunto(s)
Corioamnionitis/microbiología , Edad Gestacional , Infecciones por Mycoplasma/diagnóstico , Complicaciones Infecciosas del Embarazo/microbiología , Adulto , Amniocentesis , Líquido Amniótico/química , Líquido Amniótico/microbiología , Carga Bacteriana , Corioamnionitis/diagnóstico , ADN Bacteriano/análisis , Femenino , Rotura Prematura de Membranas Fetales/microbiología , Humanos , Mediadores de Inflamación/análisis , Embarazo , Complicaciones Infecciosas del Embarazo/diagnóstico , Estudios Prospectivos , Índice de Severidad de la Enfermedad , Adulto JovenRESUMEN
OBJECTIVE: CD11b is an integrin molecule located on the surface of leukocytes. CD11b is involved in the processes of cell adhesion and migration. Expression of CD11b increases during inflammation. Therefore, this study was aimed at the evaluation of concentrations of CD11b in the amniotic fluid from pregnancies complicated by preterm prelabor rupture of the membranes (PPROM), with respect to the presence of microbial invasion of the amniotic cavity (MIAC), intra-amniotic inflammation (IAI), and microbial-associated IAI (the presence of both MIAC and IAI). METHODS: Eighty women with singleton pregnancies complicated by PPROM were included. Amniotic fluid samples were obtained by transabdominal amniocentesis. Amniotic fluid CD11b concentrations were determined by enzyme-linked immunosorbent assay. MIAC was determined by a non-cultivation approach. IAI was defined by a bedside amniotic fluid interleukin-6 concentration ≥745 pg/mL. RESULT: Women with MIAC or microbial-associated IAI had higher CD11b concentrations in the amniotic fluid than women without these complications (with MIAC: median 0.31 ng/mL versus without MIAC: median 0.17 ng/mL, p = .001; with microbial associated-IAI: median 0.35 ng/mL versus without microbial-associated IAI: median 0.16 ng/mL; p =.02). The presence of IAI was not associated with elevated CD11b concentrations. A weak negative correlation was found between amniotic fluid CD11b concentrations and interleukin-6 concentrations (rho = 0.26; p = .02). CONCLUSIONS: MIAC and microbial-associated IAI are characterized by higher amniotic fluid CD11b concentrations in pregnancies complicated by PPROM.
Asunto(s)
Corioamnionitis , Rotura Prematura de Membranas Fetales , Amniocentesis , Líquido Amniótico/metabolismo , Corioamnionitis/metabolismo , Femenino , Rotura Prematura de Membranas Fetales/metabolismo , Edad Gestacional , Humanos , Recién Nacido , EmbarazoRESUMEN
OBJECTIVE: Macrophage inflammatory protein 1α is a chemokine produced by various immune, epithelial, mesothelial, and fibroblast cells after exposure to bacterial lipopolysaccharide or pro-inflammatory molecules. The primary aim of this study was to determine MIP-1α concentrations in amniotic and cervical fluids from pregnancy with spontaneous preterm labor with intact membranes (PTL) with respect to the presence of intra-amniotic infection (both microbial invasion of the amniotic cavity and intra-amniotic inflammation) and sterile intra-amniotic inflammation (intra-amniotic inflammation alone). The secondary aim was to assess the diagnostic indices of MIP-1α in predicting intra-amniotic infection. MATERIALS AND METHODS: Seventy-four women with PTL were included in this study. Paired amniotic and cervical fluid samples were obtained using transabdominal amniocentesis and a Dacron polyester swab, respectively. Microbial invasion of the amniotic cavity was diagnosed based on a combination of culture and molecular biology methods. The concentration of IL-6 in the amniotic and cervical fluids was measured using an automated electrochemiluminescence immunoassay method. Intra-amniotic inflammation was defined as an amniotic fluid IL-6 concentration of ≥3000 pg/mL. The MIP-1α concentrations in the samples were assessed using an enzyme-linked immunosorbent assay. RESULTS: A difference in amniotic fluid MIP-1α was observed among women with intra-amniotic infection, sterile intra-amniotic inflammation, and negative amniotic fluid (infection: median 1779.0 pg/mL; sterile, median 102.7 pg/mL; negative, median 19.9 pg/mL; p < .0001). No difference in the concentrations of MIP-1α was identified in cervical fluid after adjustment for gestational age at sampling (infection: median 77.7 pg/mL, sterile: median 152.7 pg/mL, negative: median 18.0 pg/mL; p = .30). The presence of intra-amniotic infection was associated with elevated MIP-1α concentrations in amniotic fluid (presence: 1779.0 pg/mL vs. absence: 26.3 pg/mL, p < .0001, area under receiver operating characteristic curve = 0.87). CONCLUSIONS: In PTL pregnancies with the presence of intra-amniotic infection, the concentration of MIP-1α is elevated in amniotic fluid but not in cervical fluid. Amniotic fluid MIP-1α may provide a useful marker for intra-amniotic infection in women with PTL.
Asunto(s)
Corioamnionitis , Rotura Prematura de Membranas Fetales , Trabajo de Parto Prematuro , Embarazo , Recién Nacido , Femenino , Humanos , Corioamnionitis/microbiología , Interleucina-6/metabolismo , Quimiocina CCL3/metabolismo , Trabajo de Parto Prematuro/diagnóstico , Trabajo de Parto Prematuro/metabolismo , Líquido Amniótico/metabolismo , Edad Gestacional , Inflamación/metabolismo , Rotura Prematura de Membranas Fetales/metabolismoRESUMEN
INTRODUCTION: To determine the levels of granzyme A in amniotic fluid in pregnancies complicated by preterm prelabor rupture of membranes (PPROM), based on the presence of microbial invasion of the amniotic cavity (MIAC) and/or intra-amniotic inflammation (IAI). METHODS OF STUDY: A total of 166 women with singleton pregnancies complicated by PPROM were included. Amniocentesis was performed at the time of admission and assessments of MIAC (using both cultivation and non-cultivation techniques) and IAI (interleukin-6 in amniotic fluid) were performed on all subjects. Based on the presence/absence of MIAC and IAI, the women were further divided into the following subgroups: intra-amniotic infection, sterile IAI, colonization, and absence of both MIAC and IAI. Amniotic fluid granzyme A levels were assessed using ELISA. RESULTS: Women with MIAC had lower levels of granzyme A in the amniotic fluid than women without this condition (with MIAC: median 15.9 pg/mL vs. without MIAC: median 19.9 pg/mL, p = .03). Women with sterile IAI had higher amniotic fluid granzyme A levels than women with intra-amniotic infection, colonization and women with the absence of either MIAC or IAI (intra-amniotic infection: median 15.6 pg/mL; sterile IAI: median 31.8 pg/mL; colonization: median 16.9 pg/mL; absence of both MIAC and IAI: median 18.8 pg/mL; p = .02). CONCLUSIONS: The presence of sterile IAI was associated with elevated levels of granzyme A in amniotic fluid.
Asunto(s)
Corioamnionitis , Rotura Prematura de Membranas Fetales , Líquido Amniótico , Corioamnionitis/diagnóstico , Femenino , Rotura Prematura de Membranas Fetales/etiología , Edad Gestacional , Granzimas , Humanos , Recién Nacido , Inflamación/complicaciones , EmbarazoRESUMEN
INTRODUCTION: To determine the amniotic fluid glucose levels in pregnancies complicated by preterm prelabor rupture of membranes (PPROM) based on the presence of microbial invasion of the amniotic cavity and/or intra-amniotic inflammation. METHODS OF STUDY: A total of 142 women with singleton pregnancies complicated by PPROM between gestational ages 24 + 0 and 36 + 6 weeks were included. Amniocentesis was performed at the time of admission. The assessments of microbial invasion of the amniotic cavity (using both cultivation and non-cultivation techniques) and intra-amniotic inflammation (amniotic fluid interleukin-6 levels ≥ 3000 pg/mL) were performed on all the women. Based on the presence of microbial invasion of the amniotic cavity and/or intra-amniotic inflammation, the women were further categorized into the subgroups: (i) intra-amniotic infection (the presence of both microbial invasion of the amniotic cavity and intra-amniotic inflammation); (ii) sterile intra-amniotic inflammation (the presence of intra-amniotic inflammation without microbial invasion of the amniotic cavity); (iii) colonization (the presence of microbial invasion of the amniotic cavity without intra-amniotic inflammation); and (iv) negative amniotic fluid (the absence of either microbial invasion of the amniotic cavity or intra-amniotic inflammation). Amniotic fluid glucose levels were assessed using enzymatic reference method with hexokinase. RESULTS: There was a difference in the amniotic fluid glucose levels among the women with intra-amniotic infection, sterile intra-amniotic inflammation, colonization, and those with negative amniotic fluid (p < .0001). No difference was found in the amniotic fluid glucose levels between women with intra-amniotic infection and those with sterile intra-amniotic inflammation [infection: median 11.6 mg/dL (0.7 mmol/L) vs. sterile: median 6.3 mg/dL (0.4 mmol/L); p = .41] and between women with colonization and negative amniotic fluid [colonization: median 21.6 mg/dL (1.2 mmol/L) vs. negative: median 23.4 mg/dL (1.3 mmol/L; p = .67]. Women with intra-amniotic infection and sterile intra-amniotic inflammation had lower amniotic fluid glucose levels than women with colonization and with negative amniotic fluid in crude analysis as well as after adjustment for gestational age at sampling. Amniotic fluid glucose level of 10 mg/dL (0.56 mmol/L) was the optimal concentration for the identification of intra-amniotic inflammation in women with PPROM. CONCLUSIONS: The presence of intra-amniotic inflammation was associated with lower amniotic fluid glucose levels in singleton pregnancies complicated with PPROM. An amniotic fluid glucose level of 10 mg/dL (0.56 mmol/L) was the optimal concentration for the identification of intra-amniotic inflammation in PPROM pregnancies. In the absence of better amniotic fluid markers, amniotic glucose could be used as a marker of intra-amniotic inflammation, with very good specificity in PPROM pregnancies.