Identification of diarrheagenic Escherichia coli by a new multiplex PCR assay and capillary electrophoresis
In this study, a multiplex PCR assay (mPCR) specific to nine virulence genes and an internal control was designed for the identification of five DEC pathotypes. A temperature switch PCR (TSP) strategy was used in the PCR amplification. The PCR products were detected by capillary electrophoresis. The limit of detection (LOD) of the 10-plex reaction was 5 × 103 copies/reaction for stx2 and 5 × 102 copies/reaction for the other targets. The mPCR showed very high specificity, and inclusivity and exclusivity were both 100%. When the mPCR assay was used for the detection of 221 cryopreserved diarrhea specimens, DEC col...
Source: Molecular and Cellular Probes - November 1, 2019 Category: Molecular Biology Source Type: research
Development and evaluation of a Luminex xTAG assay for sulfonamide resistance genes in Escherichia coli and Salmonella isolates
Publication date: Available online 31 October 2019Source: Molecular and Cellular ProbesAuthor(s): Fengjiao Xu, Fangui Min, Jing Wang, Yinzhu Luo, Shuwu Huang, Meiling Chen, Ruike Wu, Yu ZhangAbstractClinically occurring sulfonamide resistance in gram-negative bacteria is codified by several sul genes, mostly associated with the mobilized genetic elements named integrons, and integrons are frequently found in plasmids. There are four sul genes (sul1, sul2, sul3 and sul4) that encode resistance to sulfonamides. The aim of the present study was to develop a bead-based xTAG assay for the simultaneous detection of all four sul ...
Source: Molecular and Cellular Probes - October 31, 2019 Category: Molecular Biology Source Type: research
Detection of shrimp hemocyte iridescent virus by recombinase polymerase amplification assay
In this study, a novel real-time isothermal recombinase polymerase amplification (RPA) assay was developed using a TwistAmp exo kit for SHIV detection. First, five primers and a probe were designed for the major capsid protein gene (GenBank: KY681039.1) according to the TwistDx manual; next, the optimal primers were selected by a comparison experiment. The primers and probe were specific for SHIV and did not react with shrimp white spot syndrome virus (WSSV), shrimp infectious hypodermal and hematopoietic necrosis virus (IHHNV), shrimp enterocytozoon hepatopenaei (EHP), and macrobrachium rosenbergii nodavirus (MrNV) sample...
Source: Molecular and Cellular Probes - October 25, 2019 Category: Molecular Biology Source Type: research
Utilization of recombinase polymerase amplification method combined with lateral flow dipstick for visual detection of respiratory syncytial virus
Publication date: Available online 22 October 2019Source: Molecular and Cellular ProbesAuthor(s): Yu-zhong Xu, Du-zhi Fang, Fang-fang Chen, Qin-fei Zhao, Chao-ming Cai, Ming-gang ChengAbstractRespiratory syncytial virus (RSV) is a major causative agent of respiratory tract infection necessitating hospitalization in children. A rapid diagnostic method would facilitate early detection of RSV infection and timely implementation of special treatment. Here, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with lateral flow dipstick (LFD) was evaluated for rapid visual detection of RSV. The pr...
Source: Molecular and Cellular Probes - October 23, 2019 Category: Molecular Biology Source Type: research
Simultaneous detection of porcine reproductive and respiratory syndrome virus and porcine circovirus 3 by SYBR Green І-based duplex real-time PCR
Publication date: Available online 23 October 2019Source: Molecular and Cellular ProbesAuthor(s): Lan-lan Zheng, Lv-ye Chai, Run-bo Tian, Yu Zhao, Hong-Ying Chen, Zhen-ya WangAbstractThe SYBR Green І-based duplex real-time PCR assay was developed for simultaneous detection of porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 3 (PCV-3) genomes. PRRSV and PCV-3 were distinguished in the same sample by their distinctive melting temperature (Tm) which was 84 °C for PRRSV and 81.5 °C for PCV-3, and other non-targeted swine viruses showed no specific melting peaks. The detection limits of ...
Source: Molecular and Cellular Probes - October 23, 2019 Category: Molecular Biology Source Type: research
Design and construction of COX-2 specific fluorescent probes
Publication date: Available online 23 October 2019Source: Molecular and Cellular ProbesAuthor(s): Yi ChenAbstractCyclooxygenase-2 (COX-2) is an inducible enzyme that plays a key role in the inflammatory cascade through the generation of prostaglandins. Recent studies have found that COX-2 is closely related to the occurrence and development of tumors, angiogenesis and metastasis of tumors. Overexpressed COX-2 has become a predictive biomarker for progression of pre-malignant towards cancer in some tissues, which makes the detection of COX-2 of great value and clinical significance. This review highlights COX-2-specific flu...
Source: Molecular and Cellular Probes - October 23, 2019 Category: Molecular Biology Source Type: research
Ginsenoside Rb1 mitigates oxidative stress and apoptosis induced by methylglyoxal in SH-SY5Y cells via the PI3K/Akt pathway
Publication date: Available online 16 October 2019Source: Molecular and Cellular ProbesAuthor(s): Fengwei Nan, Guibo Sun, Weijie Xie, Tianyuan Ye, Xiao Sun, Ping Zhou, Xi Dong, Jiafu Sun, Mengren Zhang, Xiaobo SunAbstractDiabetic encephalopathy is a severe diabetic complication characterized by cognitive dysfunction and neuropsychiatric disability. Methylglyoxal (MGO), a highly reactive metabolite of hyperglycemia, serves as a major precursor of advanced glycation end products that play key roles in diabetic complications. Ginsenoside Rb1 (abbreviated as Rb1) has received extensive attention due to its potential therapeuti...
Source: Molecular and Cellular Probes - October 17, 2019 Category: Molecular Biology Source Type: research
miR-124 ameliorates depressive-like behavior by targeting STAT3 to regulate microglial activation
Publication date: Available online 15 October 2019Source: Molecular and Cellular ProbesAuthor(s): Danning Lou, Jun Wang, Xiaohang WangAbstractMajor depressive disorder (MDD) is one of the most prevalent psychiatric disorders worldwide and a major public health concern that is associated with grave consequences. Systemic complexity and feedback processes among diverse drivers of the depression disorder contribute to the considerable variation in responses to the treatment of depression. Dysfunctional microRNA (miRNA) is involved in MDD. miR-124 is enriched in the brain and may be critical in neuronal differentiation. Previo...
Source: Molecular and Cellular Probes - October 16, 2019 Category: Molecular Biology Source Type: research
Rapid and sensitive real-time recombinase polymerase amplification for detection of Marek's disease virus
In this study, a rapid and easy-to-use assay based on recombinase polymerase amplification (RPA) was developed for MDV detection. Primer-probe sets targeted the highly conserved region of Meq gene were designed and applied to the RPA assay. The assay was carried out on a real-time thermostatic fluorescence detector at 39 °C for 20 min. As revealed by the results, no cross-reactions were founded with the Newcastle disease virus (NDV), avian infectious anemia virus (CAV), infectious bursal disease virus (IBDV), avian infectious bronchitis virus (IBV), infectious laryngotracheitis virus (ILTV), avain influenza virus (AIV...
Source: Molecular and Cellular Probes - October 1, 2019 Category: Molecular Biology Source Type: research
Development of HyBeacon® probes for the forensic detection of Panthera, rhinoceros, and pangolin species
Publication date: Available online 26 September 2019Source: Molecular and Cellular ProbesAuthor(s): Kimberley George, Alice Masters, Nick DawnayAbstractThe Illegal Wildlife Trade (IWT) represents a multi-billion dollar black-market industry whereby wild species are illegally taken from their natural environment and sold. A common question asked by wildlife forensic scientists pertains to species and/or genus identity, which currently requires multi-step processing. Our work details the development of three HyBeacon® probes, used for the presumptive detection of rhinoceros, pangolin and key target species in the Panthera g...
Source: Molecular and Cellular Probes - September 27, 2019 Category: Molecular Biology Source Type: research
Computational selection of minimum length groESL operon required for Anaplasma species attribution and strain diversity analysis
In this study, on the basis of all or nearly complete groESL sequences available in the GenBank, we firstly selected a minimum partial length sequence which allows species delineation and gives a similar topology to that found by analyzing the complete sequence. By using other in silico analyses, we obtained two minimal partial sequences that are the most interesting to describe intra-specific diversity within A. ovis and A. centrale. Our results raise concern on the use of randomly selected partial sequences of groESL operon employed for the detection and the characterization of Anaplasma species and provide additional ba...
Source: Molecular and Cellular Probes - September 27, 2019 Category: Molecular Biology Source Type: research
Comparative evaluation of loop-mediated isothermal amplification (LAMP) assay, GeneXpert MTB/Rif and multiplex PCR for the diagnosis of tubercular lymphadenitis in HIV-infected patients of North India
ConclusionThe current study highlights the potential of LAMP test for the specific diagnosis of tubercular lymphadenitis in FNAC samples from HIV-infected patients, especially when cytology is either non-conclusive or non-available. Though MPCR had a higher sensitivity than LAMP assay, the added advantages of low cost, minimal technical expertise and simplicity of procedure make LAMP assay a suitable diagnostic test in resource-limited settings. (Source: Molecular and Cellular Probes)
Source: Molecular and Cellular Probes - September 22, 2019 Category: Molecular Biology Source Type: research
Editorial Board
Publication date: October 2019Source: Molecular and Cellular Probes, Volume 47Author(s): (Source: Molecular and Cellular Probes)
Source: Molecular and Cellular Probes - September 22, 2019 Category: Molecular Biology Source Type: research
A novel approach for detection of brucella using a real-time recombinase polymerase amplification assay
Publication date: Available online 18 September 2019Source: Molecular and Cellular ProbesAuthor(s): Lide Qin, Wenlong Nan, Yong Wang, Yueyong Zhang, Pengfei Tan, Yuqi Chen, Kairong Mao, Yiping ChenAbstractBrucella, the etiological agent of brucellosis, is an important zoonosis pathogen worldwide. Brucella infects humans and various domestic and wild animals, and represents a great threat to public health and animal husbandry. In the present study, we developed a real-time recombinase polymerase amplification (RPA) assay for the detection of Brucella. The assay targeted the bcsp31 gene of Brucella, and an RPA exo probe and ...
Source: Molecular and Cellular Probes - September 19, 2019 Category: Molecular Biology Source Type: research
Development and evaluation of a direct TaqMan qPCR assay for the rapid detection of diverse carnivore amdoparvoviruses
Publication date: Available online 12 September 2019Source: Molecular and Cellular ProbesAuthor(s): Yan-Hong Wu, Tao Wei, Xiu-Ting Zhang, Yong-Qiang Zhao, Jian-Ke Wang, Li Cong, Bao-Zeng Xu, Xi-Qun ShaoAbstractAmdoparvoviruses infect carnivore species, including mink, raccoon dog, fox, skunk, and red panda. Amdoparvovirus infection is a major cause of morbidity and mortality in farmed minks. Here, we developed a direct TaqMan qPCR assay for detection and quantification of carnivore amdoparvoviruses by using three primers and one probe based on the conserved VP2 gene. The detection limit for Aleutian mink disease virus (AMD...
Source: Molecular and Cellular Probes - September 13, 2019 Category: Molecular Biology Source Type: research
