Radka Bolehovska

The microbial load with genital mycoplasmas correlates with the degree of histologic chorioamnionitis in preterm PROM

OBJECTIVE We sought to determine whether there is an association between bacterial load of genital mycoplasmas and histologic chorioamnionitis (HCA) in women with preterm premature rupture of membranes (PPROM). STUDY DESIGN A total of 103 women with PPROM between 24-36 weeks of gestation were included in the study. Amniocenteses were performed, and the amounts of target genital mycoplasma DNA in amniotic fluid samples were evaluated using real-time polymerase chain reaction. The bacterial load of the genital mycoplasmas was relatively assessed using the threshold cycle value. RESULTS The presence of genital mycoplasmas in amniotic fluid was found in 38% (39/103) of the women. The presence of HCA was associated with lower threshold cycle values (median 21.3, interquartile range, 16.5-28.5, vs median 29.4, interquartile range, 27.0-30.5; P = .005). CONCLUSION HCA in PPROM is associated with a higher bacterial load of genital mycoplasmas.

The impact of the microbial load of genital mycoplasmas and gestational age on the intensity of intraamniotic inflammation

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.

Amniotic fluid soluble Toll-like receptor 4 in pregnancies complicated by preterm prelabor rupture of the membranes.

Objective: To determine amniotic fluid soluble Toll-like receptor 4 (sTLR4) levels in women with preterm prelabor rupture of the membranes according to the presence of microbial invasion of the amniotic cavity and histological chorioamnionitis and its relation to neonatal outcome. Methods: One hundred two women with singleton pregnancies with a gestational age between 24 + 0 and 36 + 6 weeks were included in a prospective cohort study. Amniocenteses were performed, and the concentrations of sTLR4 in the amniotic fluid were determined using sandwich enzyme-linked immunosorbent assay technique. Results: Women with the presence of microbial invasion of the amniotic cavity had higher sTLR4 levels [median 54.2 ng/mL, interquartile range (IQR) 10.15–289.9] than those without this condition (median 18.1 ng/mL, IQR 8.1–29.9; p = 0.001). Women with the presence of histological chorioamnionitis had a higher sTLR4 level (median 28.0 ng/mL, IQR 11.15–178.1) compared with women without histological chorioamnionitis (median 13.0 ng/mL, IQR 7.8–28.7; p = 0.003). A mixed linear model was used to adjust for confounders. The difference was found only between women with and without microbial invasion of the amniotic cavity in this model. Conclusions: Microbial invasion of the amniotic cavity was associated with higher amniotic fluid sTLR4 levels independent of confounders.

Leptospirosis: possibilities of early laboratory and clinical diagnosis

This retrospective study aims to identify and describe the problems associated with the laboratory and clinical diagnosis of leptospirosis. A total of 4,813 patients with suspected leptospirosis from an area of the Czech Republic, with a total population of 1.15 million, were examined during the period 2002–2010. Our study included only 855 patients: 545 men (mean age 41.03 ± 19.24) and 310 women (mean age 41.47 ± 20.3) who were examined using microscopic agglutination test (MAT) and a polymerase chain reaction (PCR). All patients and their physicians filled in questionnaires, which included anamnestic data, clinical symptoms and the results of laboratory tests. Out of total suspected, 89 patients (1.85%), tested positive for leptospirosis, of which 50 have been examined only serologically by MAT. Of 855 patients in our study undergoing both PCR and MAT tests, 39 have tested positive for leptospirosis. The most frequent symptom in patients with leptospirosis included fever (91.6%) and headache (69.4%). The correct laboratory diagnosis of leptospirosis depends on biological material being tested before the start of antibiotic treatment, since leptospires are extremely sensitive to antibiotics. Consequently, the PCR results alone may produce a false negative result after 24 hours following treatment with antibiotics.

Gastric fluid used to assess changes during the latency period in preterm prelabor rupture of membranes

ObjectiveTo 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).MethodsA 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.ResultMicroorganisms 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.ConclusionsThe 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.

Interleukin-6 measured using the automated electrochemiluminescence immunoassay method for the identification of intra-amniotic inflammation in preterm prelabor rupture of membranes

Abstract Introduction: We aimed to compare the amniotic fluid interleukin (IL)-6 concentrations measured using the automated electrochemiluminescence immunoassay method and ELISA, and to establish an IL-6 concentration cut-off value for intra-amniotic inflammation (IAI) in preterm prelabor rupture of membranes (PPROM), which can be used in the automated electrochemiluminescence immunoassay method. Materials and methods: A total of 120 women with PPROM were included in this study. Amniotic fluid samples were obtained through transabdominal amniocentesis. IL-6 concentrations were assessed using both the automated electrochemiluminescence immunoassay method and ELISA, the current gold standard. IAI was defined as an amniotic fluid IL-6 concentration of ≥2600 pg/mL measured using ELISA. Results: A correlation between both assays was found (Spearman’s rho = 0.97; p < .0001). Based on the receiver-operating characteristic curve for the identification of IAI (area under the curve = 0.99), a cut-off value of ≥3000 pg/mL was selected for the automated electrochemiluminescence immunoassay method with a sensitivity of 88%, specificity of 99%, positive predictive value of 97%, negative predictive value of 96%, and likelihood ratio of 76. Conclusions: For amniotic fluid IL-6 concentrations assessed using the automated electrochemiluminescence immunoassay method, a cut-off value of 3000 pg/mL was indicated for diagnosing IAI in women with PPROM.

Characterization of KPC-encoding plasmids from Enterobacteriaceae isolated in a Czech hospital

ABSTRACT Ten Enterobacteriaceae isolates collected in a Czech hospital carried blaKPC-positive plasmids of different sizes (∼30, ∼45, and ∼80 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.