A simple method for diagnosing M. tuberculosis infection in clinical samples using PCR.


:Species identification of Mycobacterium tuberculosis remains a cumbersome process. We have developed a simple method for treating clinical samples which permits direct polymerase chain reaction (PCR) amplification of mycobacterial target DNA without organic extraction. Samples were boiled for 30 min in TE-Triton, then were subjected to 30 cycles of amplification using primers derived from the repetitive clone pMTb4 developed by Patel and coworkers (1990; Journal of Clinical Microbiology 28, 513). Specificity of amplification was confirmed by hybridization with a specific probe labelled non-isotopically. In a model system consisting of cultured bacilli, this system routinely allows detection of a single organism in a sample. In preliminary studies examining applicability of this method to 96 clinical samples, 74 were positive by both PCR and mycobacterial culture, and eight were negative by both methods. Fourteen samples were negative by culture and positive by PCR, and none were positive by culture and negative by PCR. These results suggest that the PCR method may provide a sensitive alternative to conventional species-specific diagnostic methods, and that non-isotopic labelling can be used to detect hybridization in this assay.


Mol Cell Probes


Sritharan V,Barker RH Jr




Has Abstract


1991-10-01 00:00:00












  • A fluorescent method for detecting low-grade 11patUPD mosaicism in Beckwith-Wiedemann syndrome.

    abstract::The quantitative evaluation of mosaicism for uniparental disomy (UPD) involving a restricted chromosomal region requires the availability of a sensitive and reproducible method that is capable of detecting even a small percentage of disomic cells and avoiding false positive and false negative results. The occurrence o...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Russo S,Mencarelli M,Cavalleri F,Selicorni A,Cogliati F,Larizza L

    更新日期:2003-12-01 00:00:00

  • No difference in antibody titers against xenotropic MLV related virus in prostate cancer cases and cancer-free controls.

    abstract::New ELISA assays were developed to measure immunoreactivity for XMRV. Antibody titers were measured in a cohort of prostate cancer cases and cancer free controls from the central United States. No statistically significant differences were observed in immunoreactivity between cases and controls for either the XMRV-env...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Sabunciyan S,Mandelberg N,Rabkin CS,Yolken R,Viscidi R

    更新日期:2011-04-01 00:00:00

  • A rapid one-tube PCR method for simultaneously differentiating homozygotes and heterozygotes of the Sp1 binding site polymorphism in collagen type Ialpha1.

    abstract::Rapid detection of single-base changes is fundamental to molecular medicine. PCR amplification of specific alleles (PASA) has previously been used as a rapid method of genotyping single-nucleotide changes, but one reaction is required for each allele. This paper describes a Bidirectional PASA (Bi-PASA) method, which w...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lundberg GA

    更新日期:2007-06-01 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

  • Use of stable dye-DNA intercalating complexes to detect cystic fibrosis mutations.

    abstract::Several dyes are now known which intercalate into double-stranded DNA with fluorescence enhancement. One such is TOTO (1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-meth yl-2,3- dihydro-(benzo-1,3-thiazole)-2-methlyidene]-quinolinium tetraiodide), a homodimer of thiazole orange, whose complexes with DNA...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Axton RA,Brock DJ

    更新日期:1994-06-01 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

  • 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

  • Development of a novel internal positive control for Taqman based assays.

    abstract::Development of rapid amplification assays for the detection and identification of biological threat agents has become a focus of diagnostic efforts in recent years. The use of real-time PCR assays as diagnostic tools depends upon two critical processes. First, nucleic acid purification must provide template that is bo...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Hartman LJ,Coyne SR,Norwood DA

    更新日期:2005-02-01 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

  • New polymorphisms within the variable number tandem repeat (VNTR) 7 locus of Mycobacterium avium subsp. paratuberculosis.

    abstract::Variable number tandem repeat (VNTR) is a frequently employed typing method of Mycobacterium avium paratuberculosis (MAP) isolates. Based on whole genome sequencing in a previous study, allelic diversity at some VNTR loci seems to over- or under-estimate the actual phylogenetic variance among isolates. Interestingly, ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Fawzy A,Zschöck M,Ewers C,Eisenberg T

    更新日期:2016-06-01 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

  • Selective growth of mosaic cells in chromosomal analysis of chorionic villi by conventional karyotyping.

    abstract::The major cause of first-trimester pregnancy loss is chromosomal abnormality, which could be detected by many methods. Conventional karyotyping based on chorionic villi (CV) culture is frequently used but may have limitations due to culture failure and selective growth of cells. In this study, we aimed to investigate ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Zhang Y,Lei Q,Liu J,Lin M,Luo L,Li T,Wang Q,Zhou C

    更新日期:2020-06-01 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

  • 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

  • Development of a realtime RT-PCR assay for the rapid detection of influenza A(H2) viruses.

    abstract::Influenza and other acute respiratory infections are of great concern for public health, causing excessive morbidity and mortality throughout the world. Influenza virus A(H2N2), which caused a pandemic of so called "Asian flu" in 1957 was expelled from the human population by the new pandemic virus subtype H3N2 in 196...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Komissarov A,Fadeev A,Kosheleva A,Sintsova K,Grudinin M

    更新日期:2017-10-01 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

  • Identification of crucial regulatory relationships between long non-coding RNAs and protein-coding genes in lung squamous cell carcinoma.

    abstract:PURPOSE:This study aimed to analyze the relationships of long non-coding RNAs (lncRNAs) and protein-coding genes in lung squamous cell carcinoma (LUSC). METHODS:RNA-seq data of LUSC deposited in the TCGA database were used to identify differentially expressed protein-coding genes (DECGs) and differentially expressed l...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wu X,Ruan L,Yang Y,Mei Q

    更新日期:2016-06-01 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

  • Easy genotyping of complement C3 'slow' and 'fast' allotypes by tetra-primer amplification refractory mutation system PCR.

    abstract::Complement C3 'slow' and 'fast' allotypes are associated with immune-mediated disorders and may affect the outcome of renal transplantation. We report a tetra-primer amplification refractory mutation system PCR (T-ARMS-PCR) that provides a rapid, reproducible and cost-effective method to genotype both complement C3 's...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Peruzzi B,Serra M,Pescucci C,Sica M,Lastraioli S,Rondelli T,Pedemonte S,Risitano AM,De Angioletti M,Piccioli P,Notaro R

    更新日期:2010-12-01 00:00:00

  • Comparison of multiplex PCR, PCR-ELISA and fluorogenic 5' nuclease PCR assays for detection of plasmid-bearing virulent Yersinia enterocolitica in swine feces.

    abstract::Swine are implicated as the principal animal reservoir for plasmid-bearing Yersinia enterocolitica (YEP(+)) strains that are pathogenic to humans. To evaluate the utility of the PCR for detection of YEP(+) strains in naturally-contaminated pig feces, samples were first enriched in Irgasan ticarcillin potassium chlorat...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Bhaduri S

    更新日期:2002-06-01 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

  • 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

  • Genetic variation of the methylenetetrahydrofolate reductase and cystathionine beta-synthase genes in Korean patients with coronary artery disease and a new polymorphism in intron 7.

    abstract::We investigated polymorphisms of methylenetetrahydrofolate reductase (MTHFR) and cystathionine beta-synthase (CBS) genes in Korean patients with coronary artery disease (CAD) and identified a new polymorphism (c-->t) in intron 7 of the CBS gene using the single-strand conformation polymorphism method. No significant d...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Hong SH,Song J,Kim JQ

    更新日期:2001-04-01 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

  • Development of TaqMan RT-qPCR for the detection of type A human respiratory syncytial virus.

    abstract::The human respiratory syncytial virus is a common respiratory pathogen in children. Improved diagnosis of the virus is dependent on the development of tools for the rapid detection and estimation of the viral loads. In the current study, RT-qPCR using TaqMan hydrolysis probe based on the F gene detection was developed...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Abdel-Moneim AS,Shehab GM,Alsulaimani AA,Al-Malky MIR

    更新日期:2017-06-01 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

  • Detection of erythromycin resistant methylase gene by the polymerase chain reaction.

    abstract::A polymerase chain reaction (PCR) protocol was developed that could specifically amplify a 520-bp region of the erythromycin resistant methylase (ermC) gene sequence. The identity of the PCR-amplified 520-bp DNA was confirmed by HinCII endonuclease restriction digestion, which produced the predicted 440-bp and 80-bp D...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Nawaz MS,Khan AA,Cerniglia CE

    更新日期:1997-10-01 00:00:00

  • Comparison and evaluation of conventional RT-PCR, SYBR green I and TaqMan real-time RT-PCR assays for the detection of porcine epidemic diarrhea virus.

    abstract::Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal disease, resulting in substantial economic losses to the swine industry worldwide. In this study, three assays, namely a conventional reverse transcription-polymerase chain reaction (RT-PCR), a SYBR Green...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Zhou X,Zhang T,Song D,Huang T,Peng Q,Chen Y,Li A,Zhang F,Wu Q,Ye Y,Tang Y

    更新日期:2017-06-01 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

  • 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