Direct detection of vanA and vanB genes in clinical specimens for rapid identification of vancomycin resistant enterococci (VRE) using multiplex PCR.


:Surveillance for vancomycin resistant enterococci (VRE) by culture can be labour intensive and time consuming. We have developed a multiplex polymerase chain reaction (MPCR) which can be performed directly on the clinical specimen. The assay allows sensitive detection of enterococci with vanA - and vanB -mediated resistance to vancomycin. DNA was purified from stool and rectal specimens using the XTRAX(TM)DNA Extraction Kit (Gull Labs). Multiplex PCR amplified vanA and vanB targets were detected using a microtiter plate EIA. Two-hundred specimens were tested by routine culture and MPCR. Culture identified 44 VRE isolates and MPCR detected 38 of the 44 culture positives. Multiplex PCR detected three additional positive VRE specimens missed by culture for a sensitivity and specificity of 86.4 and 98.1%, respectively. When the presence of PCR inhibitors was addressed in the six culture positive/MPCR negative specimens, four additional VRE positive specimens were detected. Performing MPCR on the original specimens and on a 1:10 dilution of all specimens to minimize the effect of inhibitors gave a sensitivity and specificity of 95.5 and 98.1%, respectively. Multiplex PCR with confirmation by microtiter plate hybridization could be completed in 8 h compared with 24-48 h required for culture.


Mol Cell Probes


Petrich AK,Luinstra KE,Groves D,Chernesky MA,Mahony JB




Has Abstract


1999-08-01 00:00:00














  • Evaluation of an alkaline phosphatase-labeled oligonucleotide probe for detection and enumeration of vibrio spp. from shrimp hatchery environment.

    abstract::An alkaline phosphatase (AP)-labeled genus-specific oligonucleotide probe was developed to detect and enumerate vibrios in shrimp larvae and their surrounding environment. The probe was evaluated using 35 laboratory isolates of Vibrio species and 29 isolates of non-vibrio species. The probe was specific for the Vibrio...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Raghunath P,Karunasagar I,Karunasagar I

    更新日期:2007-08-01 00:00:00

  • Specific detection of Campylobacter concisus by PCR amplification of 23S rDNA areas.

    abstract::The phenotypic detection of Campylobacter concisus, a species of considerable genomic and phenotypic heterogeneity, has proven to be rather tedious in the past. Although alternative methods like DNA:DNA hybridization, immunotyping or whole-cell protein electrophoresis are valuable for the specific detection of C. conc...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bastyns K,Chapelle S,Vandamme P,Goossens H,De Wachter R

    更新日期:1995-08-01 00:00:00

  • Construction of an internal control for the detection of Chlamydia pneumoniae by PCR.

    abstract::For the detection of Chlamydia pneumoniae by polymerase chain reaction (PCR) in respiratory samples, an internal control was constructed to monitor the efficiency of amplification in each reaction. The internal control was designed in a way that the same primer pair can be used to amplify the internal control and targ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Ursi D,Ieven M,Van Bever HP,Goossens H

    更新日期:1998-08-01 00:00:00

  • Detection of PCR products using PNA strand invasion.

    abstract::The unique ability of homopyrimidine peptide nucleic acid (PNA) to strand invade homopurine sites of duplex DNA offers a potential alternative to existing techniques for rapid detection of PCR products. From gel shift studies, PNA was found to specifically strand invade homopurine sites that had been incorporated into...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Drewe LJ,Brightwell G,Hall EA

    更新日期:2000-10-01 00:00:00

  • Multiplex PCR for avian pathogenic mycoplasmas.

    abstract::Mycoplasma infections are of great concern in avian medicine, because they cause economic losses in commercial poultry production. A multiplex polymerase chain reaction (PCR) was optimized to simultaneously detect four pathogenic species of avian mycoplasmas. Four sets of oligonucleotide primers specific for Mycoplasm...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wang H,Fadl AA,Khan MI

    更新日期:1997-06-01 00:00:00

  • A pncA polymorphism to differentiate between Mycobacterium bovis and Mycobacterium tuberculosis.

    abstract::The pyrazinamidase gene coding for the enzyme that activates the bactericidal drug pyrazinamide contains a polymorphic site that is preserved in Mycobacterium bovis. We synthesized two sets of primers, one encompassing a 180 bp fragment, and the second spanning a 726 bp fragment including the full pncA gene. Following...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Barouni AS,Augusto CJ,Lopes MT,Zanini MS,Salas CE

    更新日期:2004-06-01 00:00:00

  • Label-free monitoring of DNA methyltransferase activity based on terminal deoxynucleotidyl transferase using a thioflavin T probe.

    abstract::We have developed a new methodology for fluorescence turn-on detection of DNA methyltransferase (MTase) activity based on terminal deoxynucleotidyl transferase (TdT) using a thioflavin T probe. This method is highly selective and sensitive. The fluorescence intensity was direct proportion to Dam MTase concentration in...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Ma C,Liu H,Li W,Chen H,Jin S,Wang J,Wang J

    更新日期:2016-04-01 00:00:00

  • Evaluation of the detection limits of PCR for identification of Mycoplasma pneumoniae in clinical samples.

    abstract::The detection limits of the polymerase chain reaction (PCR) for Mycoplasma pneumoniae were determined using specimens from persons known to have had M. pneumoniae pneumonia. Four primers were selected from the known sequence of the P1 gene. The primer pair (P1-178 and P1-809) which generates a 631 fragment gave the lo...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Leng Z,Kenny GE,Roberts MC

    更新日期:1994-04-01 00:00:00

  • A multiplex PCR for detection of Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in clinical specimens.

    abstract::A multiplex PCR was developed that is capable of detecting four of the most important bacterial agents of atypical pneumonia, Mycoplasma pneumoniae, Chlamydophila pneumoniae, Legionella pneumophila, and Bordetella pertussis in uncultured patient specimens. These organisms cause similar symptomologies and are often not...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: McDonough EA,Barrozo CP,Russell KL,Metzgar D

    更新日期:2005-10-01 00:00:00

  • Development of a SYBR green I-based duplex real-time fluorescence quantitative PCR assay for the simultaneous detection of porcine epidemic diarrhea virus and porcine circovirus 3.

    abstract::The development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for oth...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Han HY,Zheng HH,Zhao Y,Tian RB,Xu PL,Hou HL,Chen HY,Yang MF

    更新日期:2019-04-01 00:00:00

  • miR-410-5p promotes the development of diabetic cardiomyopathy by suppressing PIM1-induced anti-apoptosis.

    abstract::Diabetic cardiomyopathy (DCM) is a common complication of diabetes mellitus that can cause many severe symptoms, such as heart failure, arrhythmia, and sudden death. However, the molecular mechanisms underlying cardiac dysfunction in DCM remain elusive. In this study, we found that miR-410-5p was increased in the myoc...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Xia X,Liang Y,Zheng W,Lin D,Sun S

    更新日期:2020-08-01 00:00:00

  • Competitive reverse transcription/polymerase chain reaction for the quantification of p53 and mdm2 mRNA expression.

    abstract::Wild-type p53 (wtp53) is a tumour suppressor gene involved in cell cycle regulation. The mdm2 protein can complex with the p53 protein and influence its function as a regulator of cell growth. To detect and quantify wtp53 and mdm2 mRNA expression, we established the competitive reverse transcription/polymerase chain r...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Totzke G,Sachinidis A,Vetter H,Ko Y

    更新日期:1996-12-01 00:00:00

  • Down-regulation of UBA6 exacerbates brain injury by inhibiting the activation of Notch signaling pathway to promote cerebral cell apoptosis in rat acute cerebral infarction model.

    abstract::This study aimed to examine the UBA6 role in brain injury mediated by acute cerebral infarction (ACI). In order to screen potential therapeutic targets for ACI, two expression profiles, including GSE97537 and GSE97533 datasets, were downloaded from the GEO database. The Venn method to identify the common DEGs. 68 up-r...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Chen Z,Liu J,Chen Q,Su M,Lu H,Yang Y,Zhou G,Zhang X,Liu Y,Dong W,Fang Q

    更新日期:2020-10-01 00:00:00

  • Successful quantification of cytomegalovirus DNA by competitive PCR and detection with capillary electrophoresis.

    abstract::Human cytomegalovirus (HCMV) is responsible for severe infections in immunocompromised patients. Viral load has recently been identified as one of the major risk factors for subsequent development of HCMV disease. In this context, we developed a protocol allowing rapid, sensitive and precise quantification of HCMV DNA...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Poirier-Toulemonde AS,Imbert-Marcille BM,Ferré-Aubineau V,Besse B,Le Roux MG,Cantarovich D,Billaudel S

    更新日期:1997-02-01 00:00:00

  • Rapid genomic typing of BK virus directly from clinical specimens.

    abstract::A simple and rapid method, PCR-restriction enzyme analysis (PCR-RE) for BK virus (BKV) typing was developed, based on the presence of type-specific restriction enzyme sites in a 327 bp PCR-generated fragment which partially encodes the VP1 protein. The enzymes, Alu I, Xmn I and Ava II, were used to digest the PCR prod...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Jin L

    更新日期:1993-08-01 00:00:00

  • A novel technique for rapid automated genotyping of DNA polymorphisms in the mouse.

    abstract::The ability to rapidly and reliably genotype mice is an important concern. Traditional methods employ labour intensive and time consuming techniques such as test crossing, gel electrophoresis or nucleic acid hybridization. Here we show that a new molecular biology workstation, the WAVE DNA Fragment Analysis System, ca...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Kuklin A,Davis AP,Hecker KH,Gjerde DT,Taylor PD

    更新日期:1999-06-01 00:00:00

  • Two-color multiplex assay for the identification of orthopox viruses with real-time LUX- PCR.

    abstract::The LUX [Light Upon eXtension] is a real-time detection system that can be used for the detection and quantification of pathogens nucleic acids. In this study we used a universal LUX approach, a variation of the LUX detection system, for identifying Orthopoxvirus nucleic acids in real time. This approach enables the d...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Aitichou M,Javorschi S,Ibrahim MS

    更新日期:2005-10-01 00:00:00

  • Rapid detection of herpes simplex virus DNA by in situ hybridization with photobiotin-labelled double-stranded DNA probes.

    abstract::An assay for rapid detection of herpes simplex virus in infected cells is described. The assay utilizes in situ hybridization with photobiotin-labelled double-stranded DNA probes prepared from HSV-1 DNA cloned in plasmid vectors. The assay provided an alternative method for earlier detection of virus in cell cultures ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Chantratita W,Henchal EA,Yoosook C

    更新日期:1989-12-01 00:00:00

  • Selection of stable reference genes for gene expression analysis in sweet potato (Ipomoea batatas L.).

    abstract::Gene expression analysis is one of the most common and important studies in biology and biomedicine. No matter for traditional blotting analysis or currently commonly used PCR strategy, all need a stable reference gene for normalizing the gene expression. To screen and select housekeeping genes as the most stable refe...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Yu J,Su Y,Sun J,Liu J,Li Z,Zhang B

    更新日期:2020-10-01 00:00:00

  • Rotavirus gene detection with biotinylated single-stranded RNA probes.

    abstract::Biotinylated single-stranded RNA probes from two of the eleven genome segments of the simian rotavirus SA11 were synthesized from cloned DNA and used in dot-blot and Northern-blot hybridization assays. Different types of membranes and conditions to prepare and use synthetic non-radioactive transcript probes were evalu...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bellinzoni R,Xi JA,Tanaka TN,Scodeller E,Estes MK

    更新日期:1989-09-01 00:00:00

  • PTPN14 acts as a candidate tumor suppressor in prostate cancer and inhibits cell proliferation and invasion through modulating LATS1/YAP signaling.

    abstract::Protein tyrosine phosphatase, non-receptor type 14 (PTPN14) exerts a profound effect in the progression of multiple malignant tumors. However, whether PTPN14 plays a role in prostate cancer has not been well investigated. Herein, we evaluated the function and potential underlying mechanism of PTPN14 in prostate cancer...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wang R,Du Y,Shang J,Dang X,Niu G

    更新日期:2020-10-01 00:00:00

  • Optimized linkage and quenching strategies for quantum dot molecular beacons.

    abstract::Quantum dot (QD) molecular beacons were explored for sequence-specific DNA detection. The effectiveness of multiple linkage strategies and fluorescence quenchers were compared in hybridization-based assays. To compare linkage strategies, covalent amide linkage and streptavidin-biotin binding were used to link semicond...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Cady NC,Strickland AD,Batt CA

    更新日期:2007-04-01 00:00:00

  • A novel peptide-based recognition probe for the sensitive detection of CD44 on breast cancer stem cells.

    abstract::Metastasis and recurrence of breast cancer remain significant clinical problems. The expression level of CD44 protein is higher in breast cancer-initiating cancer stem cells; therefore, the early detection of CD44 using a sensitive diagnostic probe is important for breast cancer diagnosis and therapeutic purposes. In ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Cho JH,Lee SC,Ha NR,Lee SJ,Yoon MY

    更新日期:2015-12-01 00:00:00

  • In situ hybridization.

    abstract::In situ hybridization is the hybridization-mediated detection of specific nucleic acid sequences within structurally intact cells or tissues. As such it uniquely provides localization of nucleic acid superimposed on observable cellular and subcellular structural detail, allowing analysis unobtainable by other hybridiz...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审


    authors: Moench TR

    更新日期:1987-09-01 00:00:00

  • Real-time PCR using SYBR Green for the detection of Shigella spp. in food and stool samples.

    abstract::Shigella spp are exquisitely fastidious Gram negative organisms that frequently get missed in the detection by traditional culture methods. For this reason, this work has adapted a classical PCR for detection of Shigella in food and stool specimens to real-time PCR using the SYBR Green format. This method follows a me...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Mokhtari W,Nsaibia S,Gharbi A,Aouni M

    更新日期:2013-02-01 00:00:00

  • Direct identification of Pseudomonas aeruginosa from blood culture bottles by PCR-enzyme linked immunosorbent assay using oprI gene specific primers.

    abstract::A PCR-enzyme-linked immunosorbent assay (PCR-ELISA) was developed for direct identification of Pseudomonas aeruginosa from positive BACTEC blood culture bottles. PCR primers were designed to target a 249 bp sequence of the oprI gene in P. aeruginosa. Biotin-labeled probe (PA3) targeted to the species-specific motif we...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Kurupati P,Kumarasinghe G,Laa Poh C

    更新日期:2005-12-01 00:00:00

  • Use of a nested PCR method for the detection of astrovirus serotype 1 in human faecal material.

    abstract::In this paper we describe a reverse-transcription nested polymerase chain reaction method for detecting human astrovirus serotype 1. It has been evaluated on 56 UK diarrhoeal stool specimens and six non-UK specimens. The method has greater sensitivity than electron microscopy and may be a useful test in areas such as ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Shi M,Sikotra S,Lee T,Kurtz JB,Getty B,Hart CA,Myint SH

    更新日期:1994-12-01 00:00:00

  • Characterization of Enterobius vermicularis in a human population, employing a molecular-based method from adhesive tape samples.

    abstract::Human infection with the parasitic nematode Enterobius vermicularis occurs worldwide, particularly in children. Although its prevalence may exceed 35% in some parts of the world, molecular studies of E. vermicularis in humans are limited. The aim of the present study was to investigate the genetic variation within E. ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Piperaki ET,Spanakos G,Patsantara G,Vassalou E,Vakalis N,Tsakris A

    更新日期:2011-04-01 00:00:00

  • Recombinant VirB5 protein as a potential serological marker for the diagnosis of bovine brucellosis.

    abstract::The molecular tag vaccine against Brucella abortus and serological testing are the main methods of prevention of brucellosis used currently. They can discriminate vaccinated animals and humans from those naturally infected. In this study, we constructed a gene deletion mutant strain, B. abortus S19 virB5 with a molecu...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Tan W,Wang XR,Nie Y,Wang C,Cheng LQ,Wang XC,Zhang R,Yan GM

    更新日期:2012-06-01 00:00:00

  • Comparison of enzyme-linked immunosorbent assay (ELISA) with dot hybridization using 32P- or 2-acetylaminofluorene (AAF)-labelled cDNA probes for the detection and characterization of beet necrotic yellow vein virus.

    abstract::Beet Necrotic Yellow Vein Virus (BNYVV) was detected by enzyme-linked immunosorbent assay (ELISA) and RNA/DNA dot hybridization using either radiolabelled or non-radioactive probes. Dot hybridization specifically distinguished isolates that could not be distinguished by ELISA. The detection thresholds for ELISA, hybri...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Sakamoto H,Lemaire O,Merdinoglu D,Guesdon JL

    更新日期:1989-06-01 00:00:00