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Vo Van Giau, Thuy Trang Nguyen, Thi Kim Oanh Nguyen, Thi Thuy Hang Le, Tien Dung Nguyen
Shiga toxin-producing Escherichia coli O157:H7 (E. coli O157:H7) strains are foodborne infectious agents that cause a number of life-threatening diseases, including hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). Shiga toxin 1 (stx1), shiga toxin 2 (stx2), or a combination of both are responsible for most clinical symptoms of these diseases. Hence, various diagnostic methods have been developed so far to detect shiga toxins such as cell culture, ELISA, Rapid Latex Agglutination (RPLA) and hybridization, but due to high costs and labor time in addition to low sensitivity, they have not received much attention...
June 2016: 3 Biotech
Kusum Sharma, Rajesh Kumar Meena, Ashish Aggarwal, Rajesh Chhabra
BACKGROUND: Establishment of a reliable and rapid diagnosis is of paramount importance in spinal tuberculosis. The available gadgetry of investigations, such as AFB smear, culture of Mycobacterium tuberculosis, and Uniplex PCR, suffers from a lack of adequate sensitivity and/or a lack of rapidity. Therefore, many times a diagnosis is made either very late in the disease process or sometimes empirical therapy has to be started because a definite diagnosis could not be made. All of these are not ideal situations for a clinician...
March 2017: Acta Neurochirurgica
Lina Ma, Liyan Wang, Yuefeng Chu, Xuerui Li, Yujun Cui, Shengli Chen, Jianhua Zhou, Chunling Li, Zhongxin Lu, Jixing Liu, Yongsheng Liu
Haemophilus parasuis is classified mainly through serotyping, but traditional serotyping always yields non-typable (NT) strains and unreliable results via cross-reactions. Here, we surveyed the serotype prevalence of Chinese H. parasuis isolates using traditional serotyping (gel immuno-diffusion test, GID) and molecular serotyping (multiplex PCR, mPCR). We also investigated why discrepant results between these methods were obtained, and investigated mPCR failure through whole-genome sequencing. Of the 100 isolate tested, 73 (73%) and 93 (93%) were serotyped by the GID test and mPCR, respectively, with a concordance rate of 66% (66/100)...
2016: PloS One
Ke-Min Zhang, Guo-Yan Zhao, Bin-Bin Zhang, Qi Xu, Chun-Ping Chu, Hua Jin, De-Lai Qiu
We recently found that extracellular administration of nicotine indirectly excited hypothalamic paraventricular nucleus (PVN) corticotropin-releasing hormone (CRH) mRNA-expressing neurons. In this study, we studied the effect of nicotine on PVN oxytocin (OT) mRNA-expressing neuron in vitro in rats, by whole-cell patch-clamp recording technique, immunohistochemistry methods and single-cell reverse-transcription multiplex polymerase chain reaction (SC-RT-mPCR) methods Our results showed that 79.3% (73/92) of the 92 PVN putative magnocellular neurons co-expressed GAPDH mRNA and OT mRNA...
December 5, 2016: Neuroscience Letters
Najmiyatul Fadilah, Alfizah Hanafiah, Hamizah Razlan, Zin Qin Wong, Isa Mohamed Rose, Md Mostafizur Rahman
BACKGROUND: No gold standard has yet been established for the diagnosis of H. pylori infection. A multiplex polymerase chain reaction (mPCR) was developed in this study for rapid, sensitive and specific detection of H. pylori from gastric biopsies. METHODS: H. pylori infections were determined by in-house rapid urease test (iRUT), culture, histology and multiplex PCR. RESULTS: A total of 140 (60.9%) from 230 patients were positive for H. pylori infection...
October 2016: British Journal of Biomedical Science
Ke Feng, Wenzhong Hu, Aili Jiang, Sarengaowa, Yongping Xu, Yu Zou, Liu Yang, Xin Wang
Fresh-cut cantaloupe is particularly susceptible to contamination with pathogenic bacteria, such as Escherichia coli O157:H7, Listeria monocytogenes, and Staphylococcus aureus. Therefore, development of rapid, yet accurate detection techniques is necessary to ensure food safety. In this study, a multiplex PCR system and propidium monoazide (PMA) concentration were optimized to detect all viable pathogens in a single tube. A dual filtration system utilized a filtration membrane with different pore sizes to enrich pathogens found on fresh-cut cantaloupe...
2016: PloS One
Guillaume Voiriot, Benoit Visseaux, Johana Cohen, Liem Binh Luong Nguyen, Mathilde Neuville, Caroline Morbieu, Charles Burdet, Aguila Radjou, François-Xavier Lescure, Roland Smonig, Laurence Armand-Lefèvre, Bruno Mourvillier, Yazdan Yazdanpanah, Jean-Francois Soubirou, Stephane Ruckly, Nadhira Houhou-Fidouh, Jean-François Timsit
BACKGROUND: Multiplex polymerase chain reaction (mPCR) enables recovery of viruses from airways of patients with community-acquired pneumonia (CAP), although their clinical impact remains uncertain. METHODS: Among consecutive adult patients who had undergone a mPCR within 72 hours following their admission to one intensive care unit (ICU), we retrospectively included those with a final diagnosis of CAP. Four etiology groups were clustered: bacterial, viral, mixed (viral-bacterial) and no etiology...
October 25, 2016: Critical Care: the Official Journal of the Critical Care Forum
Abazar Pournajaf, Ramazan Rajabnia, Mansour Sedighi, Aziz Kassani, Vahid Moqarabzadeh, Lida Lotfollahi, Abdollah Ardebilli, Behzad Emadi, Gholamreza Irajian
INTRODUCTION: This study aimed to determine the prevalence, and virulence factors of Listeria monocytogenes isolated from various samples by multiplex polymerase chain reaction (MPCR). METHODS: A total of 617 isolates were obtained and MPCR was employed for detection of the inlA, inlC, and inlJ genes. RESULTS: L. monocytogenes was detected in 46 (7.45%) of the 617 specimens. inlA, inlC, and inlJ were detected in 100%, 76.26%, and 71% isolates, respectively...
September 2016: Revista da Sociedade Brasileira de Medicina Tropical
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...
November 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
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