Non-radioactive detection of Mycobacterium tuberculosis LCR products in a microtitre plate format.


:As part of the development of the ligase chain reaction (LCR) into a tool which can be used by a wide variety of researchers, we have investigated several analytical detection systems for the products of this amplification reaction. While early work with this technology has used gel electrophoresis to separate the LCR probes from the ligated product, solid phase capture techniques are also applicable, particularly when one of the probes is modified with a 'hook' such as biotin, and the adjoining probe modified with a detectable label. In this study we report a comparison of eight different non-radioactive detection techniques and discuss the analytical sensitivity of each. Detection with laser scanning fluorescent gel electrophoresis remains the most sensitive, with the assay described herein capable of detecting 100 molecules of the Mycobacterium tuberculosis insertion element IS6110 in a background of 4 micrograms of unrelated DNA. This method was followed closely by solid-phase capture and chemiluminescence detection which gave a sensitivity of 1000 molecules of IS6110. Fluorescence detection was approximately 10-fold less sensitive than chemiluminescence detection, and absorbance detection was a further 10-fold less sensitive than fluorescence detection. However, absorbance detection even at this level can still be useful for systems where visual interpretation is desired.


Mol Cell Probes


Winn-Deen ES,Batt CA,Wiedmann M




Has Abstract


1993-06-01 00:00:00














  • Diagnosis of mouse hepatitis virus contamination in mouse population by using nude mice and RT-PCR.

    abstract::Mouse hepatitis virus (MHV) infection in laboratory mouse populations is a serious problem, because the MHV infections are known to interfere with research results. Confirmation of indirect serological detection methods by viral isolation is difficult. Reverse transcription plus polymerase chain reaction (RT-PCR) was ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wang RF,Campbell WL,Cao WW,Colvert RM,Holland MA,Cerniglia CE

    更新日期:1999-02-01 00:00:00

  • Genomic structure of the EPHA1 receptor tyrosine kinase gene.

    abstract::Some receptor tyrosine kinase genes are mutated in inherited and somatically acquired human cancers. To permit mutational analysis, the complete genomic structure of the human EPHA1 gene on chromosome 7q34 was determined and oligonucleotide pairs were designed to amplify coding regions. The gene contains 18 exons, two...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Owshalimpur D,Kelley MJ

    更新日期:1999-06-01 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

  • DNA typing of HLA-B27 by polymerase chain reaction.

    abstract::To find a specific method for HLA-B27 typing, we tested an HLA-B27-specific polymerase chain reaction. This method was used for screening 100 randomly selected blood donors, 10 of them being HLA-B27 positive. A flow cytometric method and this PCR method were compared. ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lucotte G,Burckel A

    更新日期:1997-08-01 00:00:00

  • Amplification of three hypervariable DNA regions by polymerase chain reaction for paternity determinations: comparison with conventional methods and DNA fingerprinting.

    abstract::The present study evaluates the usefulness of a PCR-based method for routine paternity testing in 35 paternity cases. This identification method which is based on amplification of three hypervariable genetic loci, apoB, D1S80 and HLA-DQ alpha, is compared, with regard to reliability and technical feasibility, to the c...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Helminen P,Sajantila A,Johnsson V,Lukka M,Ehnholm C,Peltonen L

    更新日期:1992-02-01 00:00:00

  • Design considerations and effects of LNA in PCR primers.

    abstract::The effects of comprehensive LNA substitution in PCR primers for amplification of human genomic DNA targets are presented in this report. Previous research with LNA in other applications has shown interesting properties for molecular hybridization including enhanced specificity in allele-specific PCR. Here we systemat...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Latorra D,Arar K,Hurley JM

    更新日期:2003-10-01 00:00:00

  • Use of DNA restriction endonuclease digest and ribosomal RNA gene probe patterns to fingerprint Helicobacter pylori and Helicobacter mustelae isolated from human and animal hosts.

    abstract::Variation amongst strains of Helicobacter pylori and Helicobacter mustelae was examined by DNA restriction endonuclease digestion and rRNA gene patterns generated using a non-radioactive probe. Chromosomal DNA was extracted from 30 cultures of H. pylori from human, Rhesus monkey and pig gastric mucosa, and from three ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Morgan DD,Owen RJ

    更新日期:1990-08-01 00:00:00

  • Development and validation of a multiplex real-time PCR for detection of Clostridium chauvoei and Clostridium septicum.

    abstract::Clostridium chauvoei is the causative agent of blackleg in cattle and sheep. The clinical symptoms of this severe disease are very similar to that of malignant edema (Clostridium septicum), infections of other Clostridium species belonging to the gas edema complex, and anthrax (Bacillus anthracis). C. chauvoei and C. ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lange M,Neubauer H,Seyboldt C

    更新日期:2010-08-01 00:00:00

  • Molecular detection and identification of enteroviruses in children admitted to a university hospital in Greece.

    abstract::Although enteroviral infections occur frequently during childhood, the circulation of particular serotypes has never been studied in Greece. The objectives of the present report were molecular detection and identification of human enteroviruses in children admitted with nonspecific febrile illness or meningitis to a u...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Siafakas N,Attilakos A,Vourli S,Stefos E,Meletiadis J,Nikolaidou P,Zerva L

    更新日期:2011-10-01 00:00:00

  • Development of a multiplex PCR assay for the identification of pathogenic genes of Escherichia coli in milk and milk products.

    abstract::A multiplex PCR for the simultaneous detection of some pathogenic genes of enteropathogenic, enterotoxigenic and verocytotoxin-producing Escherichia coli was developed. In this study primers found in literature as well as primers to the purpose designed were used. In this way, it was possible to generate specific frag...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bottero MT,Dalmasso A,Soglia D,Rosati S,Decastelli L,Civera T

    更新日期:2004-08-01 00:00:00

  • Quantitative PCR evaluation of deletions/duplications identified by array CGH.

    abstract::Genomic deletions/duplications detected by array comparative genomic hybridization (aCGH) should be confirmed by an independent technology. This approach allows also to test, at low cost, inheritance of the imbalance. In the present study we explored the use of quantitative PCR (qPCR) to confirm aCGH-detected potentia...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Baldan F,Passon N,Burra S,Demori E,Russo PD,Damante G

    更新日期:2019-08-01 00:00:00

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

    abstract::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 resi...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


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

    更新日期:1999-08-01 00:00:00

  • Potential regulatory SNPs in promoters of human genes: a systematic approach.

    abstract::Single nucleotide polymorphisms (SNPs) can significantly contribute to the cellular level of the mRNA transcripts encoded by human disease related genes. DNA variations between individuals can be an indication of predisposition to disease or affect the response to treatment. An algorithm allowing in silico extraction ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Stepanova M,Tiazhelova T,Skoblov M,Baranova A

    更新日期:2006-12-01 00:00:00

  • 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.

    abstract::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 sub...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Dhungyel OP,Whittington RJ,Egerton JR

    更新日期:2002-08-01 00:00:00

  • Real-time monitoring of DNA methyltransferase activity using a hemimethylated smart probe.

    abstract::A real-time assay for DNA methyltransferase (MTase) activity has been developed. A hemimethylated smart probe is used as the substrate for DNA MTase. Cleavage of the methylated product leads to separation of fluorophore from quencher, giving a proportional increase in fluorescence. The method permits real-time monitor...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Jin S,Liu H,Xia K,Ma C,He H,Wang K

    更新日期:2016-06-01 00:00:00

  • A simple PCR-based genotyping method for M105I mutation of alpha-SNAP enhances the study of early pathological changes in hyh phenotype.

    abstract::alpha-SNAP is an essential component of the protein machinery responsible for membrane fusion events in different cell types. The hyh (hydrocephalus with hop gait) mouse carries a missense mutation in Napa gene that results in a point mutation (M105I) in alpha-SNAP protein. Homozygous animals for the mutant allele hav...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bátiz LF,Roales-Buján R,Rodríguez-Pérez LM,Matas IM,Páez P,Roque M,Jiménez AJ,Ramos C,Pérez-Fígares JM

    更新日期:2009-12-01 00:00:00

  • PVT1 induces NSCLC cell migration and invasion by regulating IL-6 via sponging miR-760.

    abstract::Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpr...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Su XH,Zhu YR,Hou YJ,Li K,Dong NH

    更新日期:2020-12-01 00:00:00

  • Specific detection of common pathogens of acute bacterial meningitis using an internally controlled tetraplex-PCR assay.

    abstract::Accurate and timely diagnosis of acute bacterial meningitis is critical for antimicrobial treatment of patients. Although PCR-based methods have been widely used for the diagnosis of acute meningitis caused by bacterial pathogens, the main disadvantage of these methods is their high cost. This disadvantage has hampere...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Farahani H,Ghaznavi-Rad E,Mondanizadeh M,MirabSamiee S,Khansarinejad B

    更新日期:2016-08-01 00:00:00

  • Development of a pair of real-time loop mediated isothermal amplification assays for detection of Yersinia pestis, the causative agent of plague.

    abstract::Yersinia pestis, the causative agent of plague mainly infects rodents, while humans are the accidental host. The conventional diagnostic methods available for Y. pestis exhibit cross-reactivity with other enteropathogenic bacteria which makes its detection difficult. Rapid and reliable point-of-care detection of Y. pe...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Singh R,Pal V,Tripathi NK,Goel AK

    更新日期:2020-12-01 00:00:00

  • Opportunities and challenges for the application of microfluidic technologies in point-of-care veterinary diagnostics.

    abstract::There is a growing need for low-cost, rapid and reliable diagnostic results in veterinary medicine. Point-of-care (POC) tests have tremendous advantages over existing laboratory-based tests, due to their intrinsic low-cost and rapidity. A considerable number of POC tests are presently available, mostly in dipstick or ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审


    authors: Busin V,Wells B,Kersaudy-Kerhoas M,Shu W,Burgess ST

    更新日期:2016-10-01 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

  • Direct extraction and molecular characterization of enteroviruses genomes from human faecal samples.

    abstract::Routine diagnosis of acute flaccid paralysis (AFP) is still based on classical virological procedures. Several enteroviruses serotypes are not easily isolated in cell cultures system used and routinely more than one passage in cell culture is performed. A total of 54 archived faecal samples were examined. The heteroge...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bolanaki E,Kottaridi C,Dedepsidis E,Kyriakopoulou Z,Pliaka V,Pratti A,Levidiotou-Stefanou S,Markoulatos P

    更新日期:2008-06-01 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

  • Specific detection of enterovirus 71 directly from clinical specimens using real-time RT-PCR hybridization probe assay.

    abstract::Enterovirus 71 (EV71) is one of the main causative agents of hand, foot and mouth disease (HFMD) in young children. Infections caused by EV71 could lead to many complications, ranging from brainstem encephalitis to pulmonary oedema, resulting in high mortality. Thus, rapid detection of the virus is required to enable ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Tan EL,Chow VT,Kumarasinghe G,Lin RT,Mackay IM,Sloots TP,Poh CL

    更新日期:2006-04-01 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

  • Development of a TB green II-based duplex real-time fluorescence quantitative PCR assay for the simultaneous detection of porcine circovirus 2 and 3.

    abstract::Porcine circovirus 3 (PCV3), as a newly emerged circovirus, is widely distributed in pig populations worldwide. Co-infection of PCV2 and PCV3 has been reported frequently in clinical samples. In the present study, a TB Green II-based duplex real-time polymerase chain reaction (qPCR) was developed to rapidly and differ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Zhao Y,Han HY,Fan L,Tian RB,Cui JT,Li JY,Chen HY,Yang MF,Zheng LL

    更新日期:2019-06-01 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

  • 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

  • The specificity of pilin DNA sequences for the detection of pathogenic Neisseria.

    abstract::A nucleic acid hybridization assay for the detection of the pilin gene of Neisseria gonorrhoeae has been devised. The method involves solution hybridization of pilin specific synthetic oligonucleotide probes to genomic DNA in crude cell lysates. This is followed by capture of the probe-target complex onto a microtitre...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Kolberg JA,Besemer DJ,Stempien MM,Urdea MS

    更新日期:1989-03-01 00:00:00

  • Pertactin-negative variants of Bordetella pertussis in New York State: a retrospective analysis, 2004-2013.

    abstract::The first report of pertactin-negative variants of Bordetella pertussis in the United States has raised questions about the role of acellular pertussis vaccines in the recent increase of pertussis cases. Our laboratory utilized a sequence-based method to identify mutations in the pertactin gene responsible for these v...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Quinlan T,Musser KA,Currenti SA,Zansky SM,Halse TA

    更新日期:2014-08-01 00:00:00