scholarly journals Evaluation of significant bacteriuria in pregnant women using quantitative real-time PCR

2016 ◽  
Vol 65 (4) ◽  
pp. 50-56
Author(s):  
Tatyana A. Khusnutdinova ◽  
Elena V Shipitsina ◽  
Yulia A. Savochkina ◽  
Olga Yu. Timoshina ◽  
Elena V. Rybina ◽  
...  

Introduction. Urinary tract infections are the most common infections in obstetrics and gynecology. Bacteriological method to investigate urine is laborious and time-consuming, therefore development of accurate and rapid methods for the detection of significant bacteriuria is important. Objective. Evaluation of quantitative real-time PCR based approach for the detection of significant bacteriuria in pregnant women. Material and methods. A retrospective investigation of mid-stream urine samples obtained from pregnant women was performed. Urine culture was performed using quantitative method, and a case was considered as significant bacteriuria if ≥ 105 CFU/ml were detected. Urine samples were analyzed for main uropathogens / groups of uropathogens using quantitative multiplex real-time PCR. Diagnostic characteristics of PCR were computed relative to the results of urine culture. Results. In total, 896 urine samples were tested. Of them, significant bacteriuria was found in 28 cases (3%). The frequency of detection of Escherichia coli was 50%, Enterococcus spp. — 25%, Klebsiella spp. — 7%, Proteus spp. and S. saprophyticus 4% each, Streptococcus spp. — 14%. Sensitivity and specificity of the detection of significant bacteriuria using quantitative real-time PCR for the majority of bacterial species / groups were 99% to 100%. Sensitivity and specificity of the quantitative real-time PCR based method were 96% and 98%, respectively. Conclusions. Prevalence of significant bacteriuria among pregnant women is 3%. Half of the uropathogens isolated from pregnant women with bacteriuria are E. coli. Sensitivity and specificity of quantitative PCR for the detection of significant bacteriuria are 96% and 98%, respectively.

Author(s):  
Arjun Bhugra ◽  
Supriya Gachinmath

Background and Objectives: Urinary tract infections (UTI) are the most common bacterial infections in both outpatient and inpatient department received for routine bacterial culture and sensitivity. We looked for significant bacteriuria in re- quested repeat urine sample after primary urine culture yielded significant growth (>105  CFU/ml) of ≥3 types of colonies. Also studied, different isolates grown with their sensitivity pattern and contamination rates of urine samples from different departments. Materials and Methods: In routine, primary urine cultures yielding ≥3 types of colonies on Cystine Lactose Electrolyte Deficient (C.L.E.D) were requested for repeat samples, collected with aseptic precautions after proper instructions. Data was analyzed for the Microbiological profile and its clinical correlation. Results: Among 617 received requested urine samples, 292 (47.3%) yielded significant bacteriuria. Clinical details were available for 252 cases out of which 100 (39.7%) showed asymptomatic bacteriuria, 87 (34.5%) complicated UTI and 65 (25.7%) uncomplicated UTI. Null hypothesis was rejected as 292 (47.3%) of the received repeat samples showed significant bacteriuria and 325 (53%) showed normal flora/no growth i.e. there is a 50% chance of getting either a positive culture or normal flora/no growth in repeat urine samples after the primary urine culture showed ≥3 types of colonies. It indicates the importance of requesting repeat urine samples for an accurate urine culture report. Male patients were significantly associ- ated with significant bacteriuria and complicated UTI (p= 0.001). Escherichia coli (n=112, 28%) was the most common fol- lowed by Klebsiella species (n=66, 16.4%) and Enterococcus species (n=69, 17.2%). 183 (45.6%) isolates were Multi-Drug Resistant (MDR) Gram Negative Bacilli (GNBs), Escherichia coli (50.3%) being most common. Vancomycin Resistant Enterococcus (VRE) (n=8, 2.0%) was also isolated. Conclusion: Our study justifies the rationale for asking a repeat urine samples which helps in providing an appropriate mi- crobiological report with antibiotic sensitivity pattern, hence preventing unwanted reporting of commensals/contaminants facilitating evidence based therapy.


2021 ◽  
Vol 45 (3) ◽  
pp. 189-192
Author(s):  
Nobuhisa Ishiguro ◽  
Hideaki Kikuta ◽  
Mutsuko Konno ◽  
Rikako Sato ◽  
Atsushi Manabe

Abstract Objectives Mycoplasma pneumoniae is one of the common causative pathogens of community-acquired respiratory tract infections mainly in children and young adults. Rapid and accurate diagnostic techniques for identifying the causative pathogen would be useful for initiating treatment with an appropriate antibiotic. The purpose of the present study was to evaluate the sensitivity and specificity of a novel immunochromatographic assay using silver amplification technology using FUJI DRI-CHEM IMMUNO AG2 and FUJI DRI-CHEM IMMUNO AG cartridge Myco (FUJIFILM Co., Tokyo, Japan) for detection of M. pneumoniae. Methods Throat swab samples were collected from 170 pediatric patients who were diagnosed with bronchitis or pneumonia. The silver amplification immunochromatographic (SAI) assay was performed using these samples and the results were compared with those of real-time PCR. The time required for the SAI assay is approximately 20 min (5 min for sample preparation and 15 min for waiting time after starting the assay). Results The sensitivity and specificity of the SAI assay for detection of M. pneumoniae were 85.2 and 99.1%, respectively, and the assay showed positive and negative predictive values of 98.1 and 92.3%, respectively, compared with the results of real-time PCR. The diagnostic accuracy was 94.1%. Conclusions FUJI DRI-CHEM IMMUNO AG2 and FUJI DRI-CHEM IMMUNO AG cartridge Myco are appropriate for clinical use. The optimal timing of this assay is five days or more after the onset of M. pneumoniae infection. However, PCR or other molecular methods are superior, especially with regard to sensitivity and negative predictive value.


2019 ◽  
Vol 12 (1) ◽  
pp. 26 ◽  
Author(s):  
Ehsan Ahmadpour ◽  
Elmira Zargami ◽  
Mahmoud Mahami-Oskouei ◽  
Adel Spotin ◽  
Abbas Shahbazi ◽  
...  

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Yi Wang ◽  
Hongjuan Liao ◽  
Yueheng Wang ◽  
Jinlin Zhou ◽  
Feng Wang ◽  
...  

Abstract Background Cardiovascular diseases have become the leading cause of death worldwide, and cardiac hypertrophy is the core mechanism underlying cardiac defect and heart failure. However, the underlying mechanisms of cardiac hypertrophy are not fully understood. Here we investigated the roles of Kallikrein 11 (KLK11) in cardiac hypertrophy. Methods Human and mouse hypertrophic heart tissues were used to determine the expression of KLK11 with quantitative real-time PCR and western blot. Mouse cardiac hypertrophy was induced by transverse aortic constriction (TAC), and cardiomyocyte hypertrophy was induced by angiotensin II. Cardiac function was analyzed by echocardiography. The signaling pathway was analyzed by western blot. Protein synthesis was monitored by the incorporation of [3H]-leucine. Gene expression was analyzed by quantitative real-time PCR. Results The mRNA and protein levels of KLK11 were upregulated in human hypertrophic hearts. We also induced cardiac hypertrophy in mice and observed the upregulation of KLK11 in hypertrophic hearts. Our in vitro experiments demonstrated that KLK11 overexpression promoted whereas KLK11 knockdown repressed cardiomyocytes hypertrophy induced by angiotensin II, as evidenced by cardiomyocyte size and the expression of hypertrophy-related fetal genes. Besides, we knocked down KLK11 expression in mouse hearts with adeno-associated virus 9. Knockdown of KLK11 in mouse hearts inhibited TAC-induced decline in fraction shortening and ejection fraction, reduced the increase in heart weight, cardiomyocyte size, and expression of hypertrophic fetal genes. We also observed that KLK11 promoted protein synthesis, the key feature of cardiomyocyte hypertrophy, by regulating the pivotal machines S6K1 and 4EBP1. Mechanism study demonstrated that KLK11 promoted the activation of AKT-mTOR signaling to promote S6K1 and 4EBP1 pathway and protein synthesis. Repression of mTOR with rapamycin blocked the effects of KLK11 on S6K1 and 4EBP1 as well as protein synthesis. Besides, rapamycin treatment blocked the roles of KLK11 in the regulation of cardiomyocyte hypertrophy. Conclusions Our findings demonstrated that KLK11 promoted cardiomyocyte hypertrophy by activating AKT-mTOR signaling to promote protein synthesis.


2008 ◽  
Vol 375 (1) ◽  
pp. 150-152 ◽  
Author(s):  
Cheng Xin Yi ◽  
Jun Zhang ◽  
Ka Man Chan ◽  
Xiao Kun Liu ◽  
Yan Hong

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