Fluorescence-based DNA minisequence analysis for detection of known single-base changes in genomic DNA.


:We describe a rapid, automated method for direct detection of known single-base changes in genomic DNA. Fluorescence-based DNA minisequence analysis is employed in a template-dependent reaction which involves a single nucleotide extension of an oligonucleotide primer by the correct fluorescently-tagged dideoxynucleotide chain terminator. Detection following electrophoresis on denaturing acrylamide gels is facilitated by alkaline phosphatase treatment of reaction products after extension followed by isopropanol precipitation of the dye-tagged, single-base-extended primer to remove unincorporated deoxynucleotides. Fluorescence analysis of the incorporated dye tag reveals the identity of the template nucleotide immediately 3' to the primer site. This technique does not require radioactivity or biotinylated PCR product, relies on the incorporation of a single dideoxynucleotide terminator to extend the primer by one nucleotide and takes advantage of the sensitivity of fluorescent terminators developed for automated DNA sequence analysis. As a demonstration, we have applied the assay to human genomic DNA for detection of the sickle mutation in the beta-globin gene, and have also examined feasibility for simultaneous delineation using a multiplex-like strategy in a single gel-lane of some of the most common beta-thalassemia mutations in the Mediterranean basin.


Mol Cell Probes


Kobayashi M,Rappaport E,Blasband A,Semeraro A,Sartore M,Surrey S,Fortina P




Has Abstract


1995-06-01 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

  • Integrated multiplex ligation dependent probe amplification (MLPA) assays for the detection of alterations in the HEXB, GM2A and SMARCAL1 genes to support the diagnosis of Morbus Sandhoff, M. Tay-Sachs variant AB and Schimke immuno-osseous dysplasia in hu

    abstract::Multiplex ligation dependent probe amplification (MLPA) assays were designed for the genes HEXB (OMIM: 606873), GM2A (OMIM: 613109) and SMARCAL1 (OMIM: 606622) of humans. Two sets of synthetic MLPA probes for these coding exons were tested. Changes in copy numbers were detected as well as single nucleotide polymorphis...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Sobek AK,Evers C,Dekomien G

    更新日期:2013-02-01 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

  • 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

  • WDR45 mutations in Rett (-like) syndrome and developmental delay: Case report and an appraisal of the literature.

    abstract::Mutations in the WDR45 gene have been identified as causative for the only X-linked type of neurodegeneration with brain iron accumulation (NBIA), clinically characterized by global developmental delay in childhood, followed by a secondary neurological decline with parkinsonism and/or dementia in adolescence or early ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审


    authors: Hoffjan S,Ibisler A,Tschentscher A,Dekomien G,Bidinost C,Rosa AL

    更新日期:2016-02-01 00:00:00

  • Detection and molecular typing of Campylobacter jejuni in fecal samples by polymerase chain reaction.

    abstract::In order to determine whether polymerase chain reaction (PCR) could be used to detect Campylobacter jejuni directly in stool samples, DNA from 66 frozen culture positive and negative fecal samples was purified by column chromatography. The flaA gene was amplified using primers directed against the conserved 5' and 3' ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Waegel A,Nachamkin I

    更新日期:1996-04-01 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

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

    abstract::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...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Winn-Deen ES,Batt CA,Wiedmann M

    更新日期:1993-06-01 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

  • 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

  • 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 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

  • Direct detection of Actinobacillus pleuropneumoniae in swine lungs and tonsils by real-time recombinase polymerase amplification assay.

    abstract::Actinobacillus pleuropneumoniae is the etiological agent of swine contagious pleuropneumoniae, which is distributed globally and associated with severe economic losses in the pig rearing industry. In this study, a real-time recombinase polymerase amplification assay (real-time RPA) based on the apxIVA gene was develop...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Li R,Wang J,Liu L,Zhang R,Hao X,Han Q,Wang J,Yuan W

    更新日期:2019-06-01 00:00:00

  • Detection of adenovirus in the waters of the Seine River estuary by nested-PCR.

    abstract::Several systems for isolating viruses from environmental samples have been tested. The most promising method is based on genomic amplification. The authors attempted to detect adenovirus in nucleic-acid extracts from the Seine River estuary by a two-step amplification of a 220-bp segment of the conserved coding region...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Castignolles N,Petit F,Mendel I,Simon L,Cattolico L,Buffet-Janvresse C

    更新日期:1998-06-01 00:00:00

  • Molecular diagnosis of genital HPV DNA types by polymerase chain reaction and sensitivity-standardized filter in situ hybridization in randomly sampled cohorts of Singapore women.

    abstract::Infection of the cervix uteri with various types of human papillomaviruses is generally considered a necessary factor in the etiology of cancer of the cervix uteri. In many human populations throughout the world, approximately 90% of cervical carcinomas are found to harbour HPV genomes, as judged by Southern blot hybr...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Chow V,Tham KM,Yeo-Gloss M,Lim-Tan SK,Sng I,Thirumoorthy T,Bernard HU

    更新日期:1990-04-01 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

  • Acquisition of uncharacterized sequences from Candidatus liberibacter, an unculturable bacterium, using an improved genomic walking method.

    abstract::An effective PCR-based genomic walking approach is described to discover previously unknown flanking genomic DNA sequences from Candidatus Liberibacter asiaticus, an unculturable, phloem-limited bacterium. Using this technique, 8564bp of new DNA sequences were obtained from three genomic loci; tufB-secE-nusG-rplKAJL-r...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lin H,Doddapaneni H,Bai X,Yao J,Zhao X,Civerolo EL

    更新日期:2008-02-01 00:00:00

  • Class D and E tetracycline resistance determinants in gram-negative bacteria from catfish ponds.

    abstract::DNA probes were used to examine tetracycline-resistant Gram-negative bacteria (281 strains representing eight species) from catfish ponds. The isolates, which did not previously hybridize with the Tet A, B and C determinants, were examined for the presence of tetracycline-resistance Tet D and Tet E determinants. The d...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: DePaola A,Roberts MC

    更新日期:1995-10-01 00:00:00

  • Multiplex RT-PCR assay for simultaneous detection of six viruses of penaeid shrimp.

    abstract::In the present study, multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developed for simultaneously detection of six major shrimp viruses including yellow-head virus (YHV), white spot syndrome virus (WSSV), Taura syndrome virus (TSV), hepatopancreatic parvovirus (HPV), infectious hypodermal and ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Khawsak P,Deesukon W,Chaivisuthangkura P,Sukhumsirichart W

    更新日期:2008-06-01 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

  • Development of PCR method specific for Marek's disease virus.

    abstract::A rapid polymerase chain reaction (PCR) assay specific for Marek's Disease Virus (MDV) was developed. This assay was able to detect MDV in inoculated chick kidney cells at dilutions of 10(-5). Negative PCR results were obtained using uninoculated chick cells, Marek's Disease Vaccine (SB), Herpesvirus of Turkeys (HVT) ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Rong-Fu W,Beasley JN,Cao WW,Slavik MF,Johnson MG

    更新日期:1993-04-01 00:00:00

  • The use of nested RT-PCR of prostate-specific membrane antigen in blood cells: implications for the detection of haematogenous neoplastic cells in patients with prostate adenocarcinoma.

    abstract::The aim of this study was to determine the presence of haematogenous neoplastic cells in patients with prostate cancer. Circulating prostate cells can be detected in cancer patients by using a nested-reverse transcriptase-polymerase chain reaction assay (RT-PCR), for prostate-specific membrane (PSM) antigen mRNA. This...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Lucotte G,Mercier G,Burckel A

    更新日期:1998-12-01 00:00:00

  • Twelve hour real-time PCR technique for the sensitive and specific detection of Salmonella in raw and ready-to-eat meat products.

    abstract::Rapid pathogen testing is vital to the food industry. Enzyme immunoassays (EIA) provide reliable negative results in 48 h, but a presumptive positive (suspect) EIA result must be confirmed by traditional culture methods, requiring an additional 72 h. Polymerase chain reaction (PCR) testing technology is accepted as an...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Ellingson JL,Anderson JL,Carlson SA,Sharma VK

    更新日期:2004-02-01 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

  • 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

  • Molecular diagnostic tests to predict the risk of progressive multifocal leukoencephalopathy in natalizumab-treated multiple sclerosis patients.

    abstract::Natalizumab is a humanized monoclonal antibody against the alpha4 chain of the alpha4beta1 and alpha4beta7 integrin heterodimers used with high effectiveness in the treatment of multiple sclerosis. The use of this drug can unfortunately be associated with the onset of progressive multifocal leukoencephalopathy, a poss...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审


    authors: Rossi F,Newsome SD,Viscidi R

    更新日期:2015-02-01 00:00:00

  • Microduplication of 10q26.3 in a Chinese hypertriglyceridemia patient.

    abstract::Hypertriglyceridemia (HTG) plays an important role in the development and progression of atherosclerosis. It is inherited in an autosomal dominant pattern with a frequency of approximately 1:1,000,000 worldwide. Previous study has demonstrated that more than six genes underlie this disorder. In addition, copy number v...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Li JJ,Chen YQ,Fan LL,Jin JY,Guo S,Xiang R

    更新日期:2018-02-01 00:00:00

  • Human papillomavirus type 33 DNA and E6-E7 transcripts in late passages of the UT-DEC-1 vaginal keratinocyte cell line.

    abstract::Transcription of human papillomavirus (HPV) type 33 early region was analysed in the UT-DEC-1 keratinocyte cell line, which has been derived from a HPV-33-containing mild vaginal dysplasia. Fifteen cDNA clones from transcripts from the E6-E7 open reading frames were constructed and analysed. Most clones represented vi...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Auvinen E,Hietanen S,Gissmann L,Alonso A

    更新日期:1998-02-01 00:00:00

  • Quantitative real-time PCR assay for Clostridium septicum in poultry gangrenous dermatitis associated samples.

    abstract::Clostridium septicum is a spore-forming anaerobe frequently implicated in cases of gangrenous dermatitis (GD) and other spontaneously occurring myonecrotic infections of poultry. Although C. septicum is readily cultured from diseased tissues it can be difficult to enumerate due to its tendency to swarm over the surfac...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Neumann AP,Dunham SM,Rehberger TG,Siragusa GR

    更新日期:2010-08-01 00:00:00

  • Multiple paternity and sperm storage in captive Hermann's tortoises, Testudo hermanni boettgeri determined from amniotic fluid adhering to the eggshell.

    abstract::We identified multiple paternity in 52.9% of the clutches of Hermann's tortoise Testudo hermanni boettgeri using polymorphic microsatellite markers. In addition we demonstrated sperm storage across seasons. DNA was extracted from the amniotic fluid adhering to the eggshell's inner surface, a procedure suitable for eas...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Farke CM,Olek K,Gerding WM,Distler C

    更新日期:2015-08-01 00:00:00