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Alexander A Firsov, Maria V Golikova, Elena N Strukova, Yury A Portnoy, Svetlana A Dovzhenko, Mikhail B Kobrin, Stephen H Zinner
To explore if combinations of linezolid (L) with rifampicin (R) are able to restrict Staphylococcus aureus resistance, the enrichment of L- and R-resistant mutants was studied in an in vitro dynamic model. L- and R-resistant mutants were enriched in all single drug treatments. In contrast, L-resistant mutants were not enriched and R-resistant mutants were similar to baseline amounts with only minimal regrowth at the end of the combination treatments. These effects appear to be explained by lowering the mutant prevention concentration (MPC) for L+R combinations (MPCL+R) compared to the MPCs of L and R alone (MPCL and MPCR) and thereby the longer times above MPCL+R (73-100% of the dosing interval for L and 42-58% for R) compared to the times above MPCL (0-44%) and MPCR (0%)...
October 17, 2016: Journal of Chemotherapy
Singh Parul, Basanti Bist, Barkha Sharma, Udit Jain, Janardan K Yadav
AIM: The present study was conducted to find the association among virulence determinants of verotoxic Escherichia coli (VTEC) isolated from cattle calf feces. MATERIALS AND METHODS: A total of 216 cattle calf fecal samples were collected aseptically and processed under required conditions for the isolation of E. coli. The isolates were further subjected to multiplex polymerase chain reaction (mPCR) for the detection of virulent genes. All the VTEC isolates were serotyped at the Central Research Institute, Kasauli, Himachal Pradesh...
August 2016: Veterinary World
Swarup A V Shah, Altaf A Kondkar, Tester F Ashavaid
BACKGROUND: With an increase in the discovery of newer genetic loci/polymorphisms in complex multifactorial diseases, there is also an increased need for methods that can simultaneously genotype multiple loci in a cost-effective manner. Using coronary artery disease (CAD) as a model, the study aimed to develop an in-house multilocus assay for simultaneous detection of 17 genetic variants in 11 genes implicated in CAD. METHODS: A multiplex polymerase chain reaction (PCR)-based reverse line blot hybridization (MPCR-RLBH) approach was used, where each DNA sample was amplified using two separate MPCRs, and the alleles were genotyped using covalently immobilized, amino-linked sequence-specific oligonucleotide probes using an enhanced chemiluminescence system...
November 2016: Scandinavian Journal of Clinical and Laboratory Investigation
Hongjun Wang, Huiling Sun, Patrick J Blackall, Zhenhua Zhang, Hongzhuan Zhou, Fuzhou Xu, Xiaoling Chen
A multiplex (m)PCR and a PCR followed by restriction fragment length polymorphism (RFLP) analysis of Avibacterium paragallinarum have been proposed as alternatives to conventional serotyping by the Page scheme. We evaluated both methods, and also sequenced the PCR-RFLP target fragment to reexamine the capacity of molecular serotyping. Eleven reference strains and 27 field isolates were used. Many reference strains and isolates were misidentified as Page serogroup B. The sequence analysis revealed 6 profiles based on the matching rates of the target sequence with the 3 reverse primers of the mPCR...
September 2016: Journal of Veterinary Diagnostic Investigation
Mitra Motallebi, Fereshteh Jabalameli, Kheirollah Asadollahi, Morovat Taherikalani, Mohammad Emaneini
The emergence of antibiotic-resistant Staphylococcus aureus in particular methicillin-resistant S. aureus (MRSA) is an important concern in burn medical centers either in Iran or worldwide. A total of 128 S. aureus isolates were collected from wound infection of burn patients during June 2013 to June 2014. Multiplex-polymerase chain reaction (MPCR) assay was performed for the characterization of the staphylococcal cassette chromosome mec (SCCmec). Genes encoding virulence factors and biofilm were targeted by PCR...
August 2016: Microbial Pathogenesis
Ram Prasnna Meena, V K Baranwal
Citrus trees harbor a large number of viral and bacterial pathogens. Citrus yellow vein clearing virus (CYVCV), Indian citrus ringspot virus (ICRSV), Citrus yellow mosaic virus (CYMV), Citrus tristeza virus (CTV) and a bacterium, Candidatus Liberibacter asiaticus (CLa) associated with huanglongbing (HLB) disease, the most prevalent pathogens in citrus orchards of different regions in India and are responsible for debilitating citriculture. For detection of these viral and bacterial pathogens a quick, sensitive and cost effective detection method is required...
September 2016: Journal of Virological Methods
Xiaowen Zheng, Gaopeng Liu, Tanja Opriessnig, Zining Wang, Zongqi Yang, Yonghou Jiang
Several novel porcine bocaviruses (PBoVs) have been identified in pigs in recent years and association of these viruses with respiratory signs or diarrhea has been suggested. In this study, an EvaGreen(®)-based multiplex real-time PCR (EG-mPCR) with melting curve analysis was developed for simultaneous detection and grouping of novel PBoVs into the same genogroups G1, G2 and G3. Each target produced a specific amplicon with a melting peak of 81.3 ± 0.34 °C for PBoV G1, 78.2 ± 0.37 °C for PBoV G2, and 85...
August 2016: Molecular and Cellular Probes
T Y Thung, N A Mahyudin, D F Basri, C W J Wan Mohamed Radzi, Y Nakaguchi, M Nishibuchi, S Radu
Salmonellosis is one of the major food-borne diseases in many countries. This study was carried out to determine the occurrence of Salmonella spp., Salmonella Enteritidis, and Salmonella Typhimurium in raw chicken meat from wet markets and hypermarkets in Selangor, as well as to determine the antibiotic susceptibility profile of S. Enteritidis and S. Typhimurium. The most probable number (MPN) in combination with multiplex polymerase chain reaction (mPCR) method was used to quantify the Salmonella spp., S. Enteritidis, and S...
August 1, 2016: Poultry Science
Ping Jin, Lijuan Wu, Shahin Oftadeh, Timothy Kudinha, Fanrong Kong, Qiyi Zeng
BACKGROUND: China is one of ten countries with the highest prevalence rate of pneumococcal infections. However, there is limited serotype surveillance data for Streptococcus pneumoniae, especially from the community or rural regions, partly due to limited serotyping capacity because Quellung serotyping is only available in few centers in China. The aim of this study was to develop a simple, practical and economic pneumococcal serotype prediction strategy suitable for future serotype surveillance in China...
2016: BMC Pediatrics
P Acosta, N Pérez, E Pérez, B Correa, C Pérez, C Gómez, V Sánchez, D G Pérez
OBJECTIVES: To evaluate the effect of dialysable leucocyte extract (DLE) on pro- and anti-inflammatory profiles in a rat model of osteoarthritis (OA). METHOD: Forty-eight male Wistar rats were divided into three groups: normal rats without treatment, OA rats treated with placebo, and OA rats treated with DLE. After treatment, the animals were killed to obtain cartilage for histological analysis and to determine the expression of pro- and anti-inflammatory cytokines by reverse transcription multiplex polymerase chain reaction (RT-MPCR) and immunohistofluorescence analyses...
April 21, 2016: Scandinavian Journal of Rheumatology
L L Liu, S J Chen, H Long, C Wang, T Y Cao, Z X Hu, D Wu
OBJECTIVE: To establish a multiplex polymerase chain reaction (mPCR) method with high sensitivity and specificity for rapid detection of common nontuberculous mycobacterium(NTM) infection in the hand. METHODS: Application of primer design software to the mycobacterium marinum, mycobacterium avium, mycobacterium kansasii and mycobacterium fortuitum, the specific gene sequences were used to design construction of multiplex PCR and detection of DNA from the non tuberculous mycobacterial standard strains of each bacterium of single PCR and multiplex DNA accuracy and sequence contrast evaluation to verify the specificity of multiple PCR primers...
April 12, 2016: Zhonghua Yi Xue za Zhi [Chinese medical journal]
Fernanda Pelisson Massi, Daniele Sartori, Larissa de Souza Ferranti, Beatriz Thie Iamanaka, Marta Hiromi Taniwaki, Maria Lucia Carneiro Vieira, Maria Helena Pelegrinelli Fungaro
We present the multiplex PCR data for the presence/absence of genes involved in OTA and FB2 biosynthesis in Aspergillus niger/Aspergillus welwitschiae strains isolated from different food substrates in Brazil. Among the 175 strains analyzed, four mPCR profiles were found: Profile 1 (17%) highlights strains harboring in their genome the pks, radH and the fum8 genes. Profile 2 (3.5%) highlights strains harboring genes involved in OTA biosynthesis i.e. radH and pks. Profile 3 (51.5%) highlights strains harboring the fum8 gene...
June 2016: Data in Brief
Jyoti Kumar, Shivendra Kumar Dixit, Rajiv Kumar
AIM: This study was aimed to detect Mannheimia haemolytica in lung tissues of sheep and from a bacterial culture. INTRODUCTION: M. haemolytica is one of the most important and well-established etiological agents of pneumonia in sheep and other ruminants throughout the world. Accurate diagnosis of M. haemolytica primarily relies on bacteriological examination, biochemical characteristics and, biotyping and serotyping of the isolates. In an effort to facilitate rapid M...
September 2015: Veterinary World
Anupama Subramony, Philip Zachariah, Ariella Krones, Susan Whittier, Lisa Saiman
OBJECTIVE: To assess whether multiplex polymerase chain reaction (mPCR) vs non-mPCR testing impacts the use of antibiotics, chest radiographs, and isolation precautions. STUDY DESIGN: We retrospectively compared use of antibiotics, chest radiographs, and isolation precautions for patients <18 years old (excluding neonates) hospitalized at a tertiary referral center tested for respiratory pathogens in the emergency department or during the first 2 hospital days, during 2 periods: June 2010-June 2012 (non-mPCR group) vs October 2012-May 2014 (mPCR group)...
June 2016: Journal of Pediatrics
F Esteves, B de Sousa, E J Calderón, L Huang, R Badura, F Maltez, Q Bassat, Y de Armas, F Antunes, O Matos
Pneumocystis jirovecii causes severe interstitial pneumonia (PcP) in immunosuppressed patients. This multicentre study assessed the distribution frequencies of epidemiologically relevant genetic markers of P. jirovecii in different geographic populations from Portugal, the USA, Spain, Cuba and Mozambique, and the relationship between the molecular data and the geographical and clinical information, based on a multifactorial approach. The high-throughput typing strategy for P. jirovecii characterization consisted of DNA pooling using quantitative real-time PCR followed by multiplex-PCR/single base extension...
June 2016: Clinical Microbiology and Infection
Xiao Liu, Wei Zhang, Xiaojing Zeng, Ruifang Zhang, Yuanping Du, Xueyu Hong, Hongzhi Cao, Zheng Su, Changxi Wang, Jinghua Wu, Chao Nie, Xun Xu, Karsten Kristiansen
High-throughput sequencing has recently been applied to profile the high diversity of antibodyome/B cell receptors (BCRs) and T cell receptors (TCRs) among immune cells. To date, Multiplex PCR (MPCR) and 5'RACE are predominately used to enrich rearranged BCRs and TCRs. Both approaches have advantages and disadvantages; however, a systematic evaluation and direct comparison of them would benefit researchers in the selection of the most suitable method. In this study, we used both pooled control plasmids and spiked-in cells to benchmark the MPCR bias...
2016: PloS One
Natacha Laprade, Michel Cloutier, David R Lapen, Edward Topp, Graham Wilkes, Richard Villemur, Izhar U H Khan
Campylobacter species are one of the leading causes of bacterial gastroenteritis in humans worldwide. This twofold study was sought to: i) develop and optimize four single-tube multiplex PCR (mPCR) assays for the detection of six virulence (ciaB, dnaJ, flaA, flaB, pldA and racR), three toxin (cdtA, cdtB and cdtC) and one antibiotic resistance tet(O) genes in thermophilic Campylobacter spp. and ii) apply and evaluate the developed mPCR assays by testing 470 previously identified C. jejuni, C. coli and C. lari isolates from agricultural water...
May 2016: Journal of Microbiological Methods
Mariana Herrera, Yudy Alexandra Aguilar, Zulma Vanessa Rueda, Carlos Muskus, Lázaro Agustín Vélez
BACKGROUND: The diagnosis of community-acquired pneumonia (CAP) caused by Legionella pneumophila, Mycoplasma pneumoniae, and Chlamydophila pneumoniae is traditionally based on cultures and serology, which have special requirements, are time-consuming, and offer delayed results that limit their clinical usefulness of these techniques. We sought to develop a multiplex PCR (mPCR) method to diagnosis these bacterial infections in CAP patients and to compare the diagnostic yields obtained from mPCR of nasopharyngeal aspirates (NPAs), nasopharyngeal swabs (NPSs), and induced sputum (IS) with those obtained with specific PCR commercial kits, paired serology, and urinary antigen...
2016: Journal of Negative Results in Biomedicine
Rujirat Hatrongjit, Anusak Kerdsin, Marcelo Gottschalk, Shigeyuki Hamada, Kazunori Oishi, Yukihiro Akeda
Multilocus sequence typing (MLST) is considered a reliable method for providing insight into the Streptococcus suis population structure, clonal relationships and the potential of particular clones to cause disease. Indeed, MLST has revealed the presence of several clonal complexes (CCs) within the Streptococcus suis population. However, the method is costly, time-consuming and difficult to use for screening large numbers of isolates. In this study, a multiplex PCR assay was developed to identify Streptococcus suis CCs that are relevant to human infections...
May 2016: Journal of Medical Microbiology
Z Cui, M R Ojaghian, Z Tao, K U Kakar, J Zeng, W Zhao, Y Duan, C M Vera Cruz, B Li, B Zhu, G Xie
AIMS: The aim of this study was to develop a multiplex PCR (mPCR) assay for rapid, sensitive and simultaneous detection of six important rice pathogens: Xanthomonas oryzae pv. oryzae, X. oryzae pv. oryzicola, Pseudomonas fuscovaginae, Burkholderia glumae, Burkholderia gladioli and Acidovorax avenae subsp. avenae. METHODS AND RESULTS: Specific primers were designed through a bioinformatics pipeline. Sensitivity of detection was established using both traditional PCR and quantitative real-time PCR on isolated DNA and on bacterial cells both in vitro and in simulated diseased seeds and the parameters were optimized for an mPCR assay...
May 2016: Journal of Applied Microbiology
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