Serogroup specific single and multiplex PCR with pre-enrichment culture and immuno-magnetic bead capture for identifying strains of D. nodosus in sheep with footrot prior to vaccination.


:The identification of Dichelobacter nodosus present in a flock is a prerequisite to specific (autogenous) vaccination. Current methods of identification of the serogroup present in a population requires that the organisms be isolated, identified visually in mixed culture on streak plates, subcultured to purify and subjected to antigenic analysis. This process takes at least 3 to 4 weeks. This study describes the development of a simple and rapid serogroup specific PCR test for D. nodosus. A common forward primer was designed from the conserved amino-terminal region of the fimbrial gene (fimA) and 9 (A-I) serogroup specific reverse primers were designed from the carboxy-terminal regions of fimA of the different serogroups. To verify the specificity within D. nodosus, each specific primer pair was tested in PCR against 18 serogroups/serotypes (prototypes) and found to be specific for all the serotypes within the homologous serogroups. Eighty four other bacterial strains, either commonly occurring in sheep or found in the environment of sheep, and including organisms related taxonomically to D. nodosus, were used to check the specificity of these assays. They were found to be specific for D. nodosus as none of the 84 bacterial stains reacted. These primers detected 1 pg of purified chromosomal DNA, or 50-100 cells of D. nodosus in crude lysates. Sensitivity was markedly improved when an immuno-magnetic capture was employed. Single tube multiplex PCRs were tested with different combinations of common forward primer and groups of 3, 4 or 5 reverse primers chosen so that amplicon size for each reaction product was different. These were able to amplify DNA of isolates from all the relevant serogroups included in the reactions. These tests were evaluated with samples taken directly from lesions of footrot, either directly or preceded by DNA purification, immuno-magnetic capture, enrichment broth culture and culture on hoof agar media. Of these methods only PCR on mixed colonies from 4-day-old cultures on 4% hoof agar media yielded results of practical value.


Mol Cell Probes


Dhungyel OP,Whittington RJ,Egerton JR




Has Abstract


2002-08-01 00:00:00














  • Multiplex PCR detection of Campylobacter jejuni and Arcobacter butzleri in food products.

    abstract::Arcobacter is a recently described species, previously considered part of the Campylobacter family. A sensitive assay such as that provided by PCR could help to distinguish the closely related Arcobacter from Campylobacter. A PCR method to specifically detect both Campylobacter jejuni and Arcobacter butzleri in the sa...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Winters DK,Slavik MF

    更新日期:2000-04-01 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

  • 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

  • A novel real-time PCR assay for highly specific detection and quantification of vaginal lactobacilli.

    abstract::PCR detection and quantification of vaginal lactobacilli remains problematic because of the high level of genetic heterogeneity and taxonomic complexity within the genus Lactobacillus. The aim of the present study was to identify conserved sequences among the genomes of major species of vaginal lactobacilli that could...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Demkin VV,Koshechkin SI,Slesarev A

    更新日期:2017-04-01 00:00:00

  • Fast and sensitive quantitative detection of HIV DNA in whole blood leucocytes by SYBR green I real-time PCR assay.

    abstract::The aim of this study was the development of a real-time PCR for HIV DNA quantification in whole blood leucocytes providing an alternative assay to those already described, almost based on the gag gene detection. The technique (pbs-rtPCR assay) is more rapid (the whole assay required less than 5h), easy to perform, om...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Casabianca A,Gori C,Orlandi C,Forbici F,Federico Perno C,Magnani M

    更新日期:2007-10-01 00:00:00

  • Transcriptional profiles of regulatory and virulence factors of Staphylococcus aureus of bovine origin: oxygen impact and strain-to-strain variations.

    abstract::Staphylococcus aureus is responsible for a large panel of infections in humans and animals. In cows, S. aureus provokes chronic intramammary infections. Little information is available about the regulation of virulence factors in bovine isolates. Moreover, oxygenation, which is low in an inflamed mammary gland, could ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Ster C,Gilbert FB,Cochard T,Poutrel B

    更新日期:2005-08-01 00:00:00

  • A species-specific DNA probe for Providencia stuartii identification.

    abstract::A DNA probe is described that can be used for identification of Providencia stuartii by means of filter hybridization assays. The probe, which is a fragment of the P. stuartii phoN gene coding for an acid phosphatase, appeared to be able to recognize only P. stuartii strains in slot-blot hybridization experiments perf...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Thaller MC,Berlutti F,Riccio ML,Rossolini GM

    更新日期:1992-10-01 00:00:00

  • A DNA microarray for identification of virulence and antimicrobial resistance genes in Salmonella serovars and Escherichia coli.

    abstract::Characterization of antimicrobial resistance and virulence gene profiles provides important information on the potential pathogenicity of bacteria. This information can be used to facilitate prompt and effective treatment of bacterial infections. We developed and tested a PCR-based microarray platform for detecting vi...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Chen S,Zhao S,McDermott PF,Schroeder CM,White DG,Meng J

    更新日期:2005-06-01 00:00:00

  • Rapid and sensitive detection of Chlamydia trachomatis using a ligatable binary RNA probe and Q beta replicase.

    abstract::A simple assay format was developed for the direct detection of C. trachomatis rRNA utilizing ligation of recombinant MDV-1 probe RNA fragments hybridized to 23S rRNA after capture and release from a solid support. Assay background (equivalent to 10(4) targets) was suppressed by blocking sequences in the 5' MDV report...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Stefano JE,Genovese L,An Q,Lu L,McCarty J,Du Y,Stefano K,Burg JL,King W,Lane DJ

    更新日期:1997-12-01 00:00:00

  • Rapid prediction of inducible clarithromycin resistance in Mycobacterium abscessus.

    abstract::We have developed a single tube TaqMan(®) real-time PCR assay that differentiates the full-length and truncated erm(41) gene to predict inducible resistance to clarithromycin in Mycobacterium abscessus. A study of 87 clinical isolates found this assay to be 90.8% concordant to conventional drug susceptibility testing ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Zhu YC,Mitchell KK,Nazarian EJ,Escuyer VE,Musser KA

    更新日期:2015-12-01 00:00:00

  • Analysis of major histocompatibility complex class I gene transcription using oligonucleotide probes.

    abstract::Many highly homologous genes are present in the murine major histocompatibility complex (MHC) class I gene family. Consequently, it is difficult to distinguish between RNA transcripts of individual class I genes solely on the basis of nucleic acid hybridization analysis using DNA probes over 50 base pairs long. To avo...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Mellor AL

    更新日期:1987-09-01 00:00:00

  • Potential of recombinant flagellin fragment from Burkholderia thailandensis as an antigen for melioidosis antibody detection by indirect ELISA.

    abstract::Non-pathogenic Burkholderia thailandensis may be used as a model for Burkholderia pseudomallei due to the genetic similarity of these species. Moreover, the experimental manipulation of B. thailandensis is safer. In this study, we constructed recombinant flagellin protein fragments of B. thailandensis E264 (FLAG300, F...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wajanarogana S,Nimnuch P,Thongmee A,Kritsiriwuthinan K

    更新日期:2013-04-01 00:00:00

  • Reverse transcription loop-mediated isothermal amplification for the rapid detection of infectious bronchitis virus in infected chicken tissues.

    abstract::A reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay targeting the nucleocapsid phosphoprotein gene of infectious bronchitis virus (IBV) was developed. The detection limits for the IBV RT-LAMP assay were 10(1) 50% egg infection dose (EID(50)) per 50 microl of titrated viruses and no cross-rea...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Chen HT,Zhang J,Ma YP,Ma LN,Ding YZ,Liu XT,Cai XP,Ma LQ,Zhang YG,Liu YS

    更新日期:2010-04-01 00:00:00

  • Enhanced discrimination of single nucleotide polymorphisms using 3' nucleotide differences in ligase detection reaction probes.

    abstract::The ligase detection reaction (LDR) is a highly specific genotyping method for single nucleotide variations. Although LDR typically discriminates single nucleotide polymorphism (SNP) alleles at the 3' end of so-called LDR discriminating probes, we designed probes in which the position of nucleotide differences for dis...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Asari M,Omura T,Maseda C,Shiono H,Tasaki Y,Matsubara K,Shimizu K

    更新日期:2010-12-01 00:00:00

  • Insulin-like growth factor-I activates NFκB and NLRP3 inflammatory signalling via ROS in cancer cells.

    abstract::Previous studies have demonstrated that insulin-like growth factor-I (IGF-1) and reactive oxygen species (ROS) are involved in the development and progression of various cancers. However, their regulatory mechanism remains unknown. In this study, we treated cancer cells (HeLa, HepG2 and SW1116 cells) and normal cells ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wang C,An Y,Wang Y,Shen K,Wang X,Luan W,Ma F,Ni L,Liu M,Yu L

    更新日期:2020-08-01 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

  • Quantification of the detection of Pneumocystis carinii by DNA amplification.

    abstract::We have developed a highly specific and sensitive technique for the detection of Pneumocystis carinii DNA using DNA amplification by the polymerase chain reaction (PCR). PCR products are detected by agarose gel electrophoresis and Southern hybridization to an oligonucleotide probe. Here we report the calibration of pa...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Peters SE,Wakefield AE,Banerji S,Hopkin JM

    更新日期:1992-04-01 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

  • Quantitative PCR for the diagnosis of cutaneous leishmaniasis from formalin-fixed and paraffin-embedded skin sections.

    abstract::The present report describes a real-time PCR-based procedure to reliably determine the quantity of Leishmania amastigotes in relation to the amount of host tissue in histological skin sections from canine and equine cases of cutaneous leishmaniasis. The novel diagnostic Leishmania-PCR has a detection limit of <0.02 am...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Müller N,Hentrich B,Frey CF,Welle M

    更新日期:2015-12-01 00:00:00

  • The construction and use of a PCR internal control.

    abstract::An example of the application and contruction of a polymerase chain reaction (PCR) internal control is presented. The internal control is synthesized in one PCR reaction. The primers used in this reaction possess 5' over-hanging ends which are identical to the primers used in the diagnostic reaction, whereas their 3' ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Sachadyn P,Kur J

    更新日期:1998-10-01 00:00:00

  • Duplex Real-time PCR assay and SYBR green I melting curve analysis for molecular identification of HPV genotypes 16, 18, 31, 35, 51 and 66.

    abstract::Long-term infection with high-risk HPV genotypes is the leading cause of cervical cancer. In the present study a Duplex Real-time PCR assay was developed in order to identify HPV types 16, 18, 31, 35, 51 and 66 in three reactions, through SYBR green I melting curve analysis. The method utilizes type-specific primer se...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Tsakogiannis D,Papacharalampous M,Toska E,Kyriakopoulou Z,Dimitriou TG,Ruether IG,Komiotis D,Markoulatos P

    更新日期:2015-02-01 00:00:00

  • Detection of shigellae, enteroinvasive and enterotoxigenic Escherichia coli using the polymerase chain reaction (PCR) in patients returning from tropical countries.

    abstract::We have used the polymerase chain reaction (PCR) to detect shigellae, EIEC and ETEC in stool specimens of diarrhoeic patients returning from tropical countries. As compared to culture (7.1% positive specimens), which recognizes only Shigella strains, PCR performed on bacterial growth from directly inoculated MacConkey...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lüscher D,Altwegg M

    更新日期:1994-08-01 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

  • SNP genotyping with FRET probes. Optimizing the resolution of heterozygotes.

    abstract::Analysis of single nucleotide polymorphisms by PCR with fluorescence resonance energy transfer (FRET) probes often can produce a result where the melting peak corresponding to perfectly matched sequence (A allele) has a smaller area than the peak corresponding to the allele with a mismatch (B allele). This imbalance c...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Martínez-García A,Sastre I,Tenorio R,Bullido MJ

    更新日期:2004-08-01 00:00:00

  • Rapid and cost effective genotyping method for polymorphisms in PPARG, PPARGC1 and TCF7L2 genes.

    abstract::Polymorphisms (rs1801282, rs8192678, rs7903146) of peroxisome proliferator-activated receptor gamma (PPARG), peroxisome proliferator-activated receptor gamma coactivator-1 alpha (PPARGC1A) and transcription factor 7-like 2 (TCF7L2) have recently been associated with different diseases, mainly type 2 diabetes. An assay...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Habalová V,Klimcáková L,Zidzik J,Tkác I

    更新日期:2009-02-01 00:00:00

  • Aptamers, the bivalent agents as probes and therapies for coronavirus infections: A systematic review.

    abstract::The recently known coronavirus, SARS-CoV-2, has turn into the greatest global health challenge, affecting a large number of societies. The lack of specific treatment and gold-standard diagnostic system has made the situation more complicated. Efforts have led to production of several diagnostic kits that are associate...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,meta分析


    authors: Torabi R,Ranjbar R,Halaji M,Heiat M

    更新日期:2020-10-01 00:00:00

  • CircHIPK3 regulates melanoma cell behaviors by binding with miR-215-5p to upregulate YY1.

    abstract:OBJECT:To investigate the role of circHIPK3 in melanoma. METHODS:Bioinformatics analysis and experiments including RT-qPCR, Pearson's correlation analysis, luciferase reporter, Western blot, and RIP assays were applied to explore the function and mechanism of circHIPK3 in melanoma. RESULTS:CircHIPK3 expression was st...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Zhu X,Sun J

    更新日期:2020-10-01 00:00:00

  • Rapid PCR using nested primers of the 16S rRNA and the hippuricase (hip O) genes to detect Campylobacter jejuni and Campylobacter coli in environmental samples.

    abstract::Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited sensitivity due to ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bang DD,Wedderkopp A,Pedersen K,Madsen M

    更新日期:2002-10-01 00:00:00

  • Tracking the extramedullary PML-RARα-positive cell reservoirs in a preclinical model: biomarker of long-term drug efficacy.

    abstract::Using an acute promyelocytic leukemia (APL) preclinical model, we show that oncogene-specific PCR (Polymerase Chain Reaction)-based assays allow to evaluate the efficacy of immunotherapy combining all-trans retinoic acid (ATRA) and a DNA-based vaccine targeting the promyelocytic leukemia-retinoic acid receptor alpha (...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Pokorna K,Le Pogam C,Chopin M,Balitrand N,Reboul M,Cassinat B,Chomienne C,Padua RA,Pla M

    更新日期:2013-02-01 00:00:00

  • A LightCycler real-time PCR hybridization probe assay for detecting food-borne thermophilic Campylobacter.

    abstract::In a previous study, we reported the performance of a PCR assay amplifying 287-bp of the 16S rRNA gene of thermo-tolerant Campylobacter (C. jejuni, C. lari, C. coli) through an international ring-trial involving 12 participating laboratories. Based on the validated set of primers, a LightCycler real-time PCR assay (LC...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Perelle S,Josefsen M,Hoorfar J,Dilasser F,Grout J,Fach P

    更新日期:2004-10-01 00:00:00