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


: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 design of sequence-specific primer sets in high identity genome sequences. The assay described here is designed to allow simultaneous detection of Variola and other orthopox viruses in a multiplex format, with a limit of detection in the range of 50--100 copies of the Orthopoxvirus genome. Regression analysis showed that the assay was linear over seven orders of magnitude, with 0.97 correlation coefficient. The sensitivity and specificity of the assay, as determined from a panel of 100 samples that contained nucleic acids from a variety of bacteria and viral species, were rated at 98%. Thus, the assay offers a sensitive and specific tool for simultaneous identification and quantification of Variola and other orthopox viruses, and the approach allows more flexible sequence-specific primers design for pox viruses as well as other microbial pathogens.


Mol Cell Probes


Aitichou M,Javorschi S,Ibrahim MS




Has Abstract


2005-10-01 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

  • Quantification of sensitive non-isotopic filter hybridizations using the peroxidase catalyzed luminol reaction.

    abstract::The development of a sensitive, non-isotopic filter hybridization method based on the peroxidase catalyzed luminol reaction is described. High sensitivity was achieved by optimizing the conditions of the hybridization procedure, the immunochemical detection and the peroxidase/luminol reaction. This resulted in the rep...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: van Gijlswijk RP,Raap AK,Tanke HJ

    更新日期:1992-06-01 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

  • 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

  • Screening South African familial adenomatous polyposis families for the five-nucleotide deletion at codon 1309 of the APC gene.

    abstract::We report the occurrence of a common five-nucleotide deletion at codon 1309 of the adenomatous polyposis coli (APC) gene in four different South African population groups. The mutation causes familial adenomatous polyposis (FAP) in 18% (4/22 unrelated patients screened) of affected South Africans, which is similar to ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Grobbelaar JJ,Oosthuizen CJ,Kotze MJ

    更新日期:1995-02-01 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

  • 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

  • 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

  • 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

  • HyBeacon probes: a new tool for DNA sequence detection and allele discrimination.

    abstract::Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleotide probe technology, termed HyBeacons, which provides a...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: French DJ,Archard CL,Brown T,McDowell DG

    更新日期:2001-12-01 00:00:00

  • Molecular contrast of EGFR expression using gold nanoparticles as a reflectance-based imaging probe.

    abstract::Advanced reflectance-based optical techniques for in vivo imaging often suffer from low contrast between neoplastic and normal tissue and are unable to image early biomolecular changes associated with carcinogenesis, thus limiting their clinical value. In this study, we exploit the resonance light scattering property ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Kah JC,Olivo MC,Lee CG,Sheppard CJ

    更新日期:2008-02-01 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

  • A method for the rapid construction of cRNA standard curves in quantitative real-time reverse transcription polymerase chain reaction.

    abstract::Quantification of nucleic acids, especially of mRNA, is increasingly important in biomedical research. The recently developed quantitative real-time polymerase chain reaction (PCR) - a highly sensitive technology for the rapid, accurate and reproducible quantification of gene expression - offers major advantages over ...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Fronhoffs S,Totzke G,Stier S,Wernert N,Rothe M,Brüning T,Koch B,Sachinidis A,Vetter H,Ko Y

    更新日期:2002-04-01 00:00:00

  • Rapid identification of Renibacterium salmoninarum using an oligonucleotide probe complementary to 16S rRNA.

    abstract::Bacterial kidney disease in salmonid fish is caused by the slow-growing Gram-positive rod, Renibacterium salmoninarum. The partial sequence of 16S rRNA from R. salmoninarum was determined and compared with published bacterial 16S rRNA sequences. From this sequence information, a 30-bases-long oligonucleotide was desig...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Mattsson JG,Gersdorf H,Jansson E,Hongslo T,Göbel UB,Johansson KE

    更新日期:1993-02-01 00:00:00

  • A multiplex RT-PCR for detection of type A influenza virus and differentiation of avian H5, H7, and H9 hemagglutinin subtypes.

    abstract::A multiplex reverse transcriptase-polymerase chain reaction (mRT-PCR) was developed and optimized for the detection of type A influenza virus; the assay simultaneously differentiates avian H5, H7 and H9 hemagglutinin subtypes. Four sets of specific oligonucleotide primers were used in this test for type A influenza vi...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Xie Z,Pang YS,Liu J,Deng X,Tang X,Sun J,Khan MI

    更新日期:2006-06-01 00:00:00

  • Up-regulated miR-374a-3p relieves lipopolysaccharides induced injury in CHON-001 cells via regulating Wingless-type MMTV integration site family member 5B.

    abstract:BACKGROUND:Osteoarthritis (OA) is a frequent and incurable joint disease, inducing significant pain and seriously threatening to human health. It has been reported that microRNAs (miRNAs) play crucial roles on cancers and inflammatory diseases via cooperating with genes. However, the effect of miR-374a-3p/Wingless-type...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Shi FL,Ren LX

    更新日期:2020-06-01 00:00:00

  • Detection of catechol-O-methyltransferase Val158Met polymorphism by a simple one-step tetra-primer amplification refractory mutation system-PCR.

    abstract::The G-->A transition at nucleotide 21881 of the human catechol-O-methyltransferase (COMT) gene represents a functional genetic polymorphism (Val158Met), rendering an enzyme with reduced activity that has been associated with psychiatric disorders and estrogen-related cancers. A new method for the detection of this pol...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Ruiz-Sanz JI,Aurrekoetxea I,Ruiz del Agua A,Ruiz-Larrea MB

    更新日期:2007-06-01 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

  • 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

  • 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

  • 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

  • Molecular cloning and nucleic acid sequencing of Chlamydia trachomatis 16S rRNA genes from patient samples lacking the cryptic plasmid.

    abstract::We have examined the relationship between Chlamydia trachomatis found in clinical samples in which the cryptic plasmid was absent and known serovars of C. trachomatis. PCR and RNase protection assays were used to compare 12 C. trachomatis serovars and a plasmidless L2 serovar strain with the reactivity of clinical spe...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: An Q,Olive DM

    更新日期:1994-10-01 00:00:00

  • Expression patterns of miR-146a and miR-146b in mastitis infected dairy cattle.

    abstract::This study reports a significant up-regulation of bta-miR-146a and bta-miR-146b expression levels in bovine mammary tissues infected with subclinical, clinical and experimental mastitis. Potential target genes are involved in multiple immunological pathways. These results suggest a regulatory function of both miRNAs f...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Wang XP,Luoreng ZM,Zan LS,Raza SH,Li F,Li N,Liu S

    更新日期:2016-10-01 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

  • Development and validation of a cost-effective in-house method, tetra-primer ARMS PCR assay, in genotyping of seven clinically important point mutations.

    abstract::The single nucleotide polymorphism (SNP) genotyping is currently considered as a particularly valuable tool for the diagnosis of different pathologies. For this reason, over the past several years a great deal of effort has been devoted to developing accurate, rapid, and cost-effective technologies for SNP analysis. A...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Etlik O,Koksal V,Arican-Baris ST,Baris I

    更新日期:2011-08-01 00:00:00

  • Experiences on the application of the polymerase chain reaction in a diagnostic laboratory.

    abstract::Double polymerase chain reaction (PCR) assays with nested primers have been applied in a routine laboratory for the diagnosis of herpes-, pesti- and retroviral infections of animals. Various methods and tools have been tested to prevent and to eliminate false positive results as well as to visualize the PCR products (...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章,评审


    authors: Belák S,Ballagi-Pordány A

    更新日期:1993-06-01 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

  • Development of PCR based assays for detection of lethal Holstein haplotype 1, 3 and 4 in Holstein Friesian cattle.

    abstract::Holstein haplotype (HH) 1, 3 and 4 are lethal mutations, responsible for early embryonic losses in Holstein Friesian (HF) cattle, worldwide. Three PCR based assays - tetra Amplification Refractory Mutation System PCR, PCR primer induced restriction analysis and PCR-restriction fragment length polymorphism techniques f...

    journal_title:Molecular and cellular probes

    pub_type: 杂志文章


    authors: Kumar A,Gupta ID,Mohan G,Vineeth MR,Ravi Kumar D,Jayakumar S,Niranjan SK

    更新日期:2020-04-01 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