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 an amplicon during the PCR cycle. This was achieved by adding a homopyrimidine sequence to the 5'-terminus of a PCR primer. The position of the strand invasion sites at the termini of the DNA duplex offers kinetic advantages for PNA strand invasion, since the termini of DNA duplexes are known to be unwound. This unwound state was demonstrated using a novel assay that determined single-stranded regions within the amplicon. The presence of the PNA moiety in strand invasion complexes was confirmed by a novel electroblot, an Enzyme Linked Nucleic Acid assay and by an increase in stability as demonstrated by T(m)studies with the Idaho RapidCycler. Since the strand invasion sites can be controlled through selection of the homopurine sequence there is great flexibility for designing strand invasion motifs unique to a particular PCR amplicon, thus providing a huge potential for differentiating and detecting multiplex PCR products.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Drewe LJ,Brightwell G,Hall EAdoi
10.1006/mcpr.2000.0311subject
Has Abstractpub_date
2000-10-01 00:00:00pages
269-83issue
5eissn
0890-8508issn
1096-1194pii
S0890850800903116journal_volume
14pub_type
杂志文章abstract：:This study evaluates five cryptic plasmid-derived DNA probes in a 4-h slot-blot hybridization assay for the detection of Chlamydia trachomatis in cultures and clinical specimens. The probes, consisting of either the entire cloned 7.5 kbp cryptic plasmid pSE8 or one of four Hin dIII/Eco RI fragments measuring 710, 1055...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/s0890-8508(05)80013-1
更新日期：1991-12-01 00:00:00
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
doi：10.1006/mcpr.1998.0201
更新日期：1998-12-01 00:00:00
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
doi：10.1016/j.mcp.2003.09.007
更新日期：2004-02-01 00:00:00
abstract：:The manual gravimetric drying moisture determination methods currently employed by most mineral processing plants fail to provide timely and accurate information required for automatic control. The costs associated with transporting and handling concentrates still represent a major portion of the overall treatment pri...
journal_title：Molecular and cellular probes
pub_type： 杂志文章,评审
doi：10.1006/mcpr.2002.0444
更新日期：2002-12-01 00:00:00
abstract：:Genetic polymorphism among patients with acute lymphoblastic leukemia (ALL) is an important factor in the effectiveness and toxicity of anti-leukemic drugs. Genotyping of various polymorphisms that impact the outcome of anti-leukemic drug therapy (pharmacogenetics) presents an attractive approach for developing indivi...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2008.02.001
更新日期：2008-06-01 00:00:00
abstract：BACKGROUND:Stroke is a leading cause of mortality worldwide. Rac-MAPK kinase 6 (Map2k6) plays important roles in cell proliferation and apoptosis. However, the role played by Map2k6 in stroke injury and the underlying mechanism of action remain unknown. METHODS:Mice received cerebral ischemia/reperfusion (I/R) injurie...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2019.101478
更新日期：2020-02-01 00:00:00
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
doi：10.1006/mcpr.1993.1027
更新日期：1993-06-01 00:00:00
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
doi：10.1016/j.mcp.2010.08.005
更新日期：2010-12-01 00:00:00
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
doi：10.1006/mcpr.1999.0228
更新日期：1999-06-01 00:00:00
abstract：:The presence of human cytomegalovirus (HCMV) was tested in 388 cervicovaginal cells specimens obtained from the same number of pregnant women. HCMV was detected in 5.41%, 11.6% and 13.9% of these specimens by conventional culture, in situ DNA hybridization and polymerase chain reaction (PCR) methods, respectively. The...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(90)90006-l
更新日期：1990-12-01 00:00:00
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
doi：10.1016/j.mcp.2016.08.004
更新日期：2016-10-01 00:00:00
abstract：:DNA aptamers (PSA-H and MT32) were applied for the detection of Apple stem pitting virus (ASPV) isolates using an Enzyme-Linked Oligonucleotide Assay (ELONA) and Western blot analysis. The specificity and effectiveness of aptamers were verified in comparison to a conventional Enzyme Linked Immunosorbent Assay (ELISA)....
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2017.10.001
更新日期：2017-12-01 00:00:00
abstract：:Angiostrongylus cantonensis, a rat lungworm, can cause eosinophilic meningitis and angiostrongyliasis in humans following ingestion of contaminated foods or intermediate/paratenic hosts with infective larvae. The snail Achatina fulica is one of the important intermediate hosts of A. cantonensis and is commonly eaten b...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2011.04.002
更新日期：2011-08-01 00:00:00
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
doi：10.1016/0890-8508(89)90037-6
更新日期：1989-03-01 00:00:00
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
doi：10.1016/j.mcp.2004.03.003
更新日期：2004-08-01 00:00:00
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
doi：10.1006/mcpr.1993.1035
更新日期：1993-06-01 00:00:00
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
doi：10.1016/s0890-8508(95)91572-9
更新日期：1995-10-01 00:00:00
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
doi：10.1016/j.mcp.2014.11.007
更新日期：2015-02-01 00:00:00
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
doi：10.1016/s0890-8508(03)00062-8
更新日期：2003-10-01 00:00:00
abstract：INTRODUCTION:Treatment in metastatic colorectal cancer (mCRC) has expanded with monoclonal antibodies targeting epidermal growth factor receptor, but is restricted to patients with a wild-type (WT) KRAS mutational status. The most sensitive assays for KRAS mutation detection in formalin-fixed paraffin embedded (FFPE) t...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2017.06.003
更新日期：2017-10-01 00:00:00
abstract：:The major outer-membrane protein (MOMP) from Campylobacter jejuni NCTC 11168 was purified by solubilization in Triton X-100. Whole-cell proteins of Campylobacter species and the MOMP were subjected to electrophoresis on sodium dodecyl sulfate-polyacrylamide gels, immunoblotting on nitrocellulose paper and enzyme-linke...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(87)90038-7
更新日期：1987-09-01 00:00:00
abstract：:Inexpensive fiberglass mesh window screens were used as spacers between colony blot filters to increase the number of bacterial isolates that could be tested by DNA colony hybridization. Sixty filters with up to 48 isolates per filter were tested at one time. This modified technique reduced the amount of radiolabelled...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(88)90009-6
更新日期：1988-12-01 00:00:00
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
doi：10.1006/mcpr.2002.0434
更新日期：2002-10-01 00:00:00
abstract：:Several studies have shown that BK viral load in plasma and urine are reliable markers for the detection of BK virus associated nephropathy (BKVAN) in renal transplant patients. We developed a quantitative real time PCR assay based on TaqMan technology for the measurement of BK viral load in plasma and urine. Consider...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2013.08.001
更新日期：2013-10-01 00:00:00
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
doi：10.1006/mcpr.1994.1040
更新日期：1994-08-01 00:00:00
abstract：:The RNA genome of pathogenic and non-pathogenic variants of citrus Hop stunt viroid (HSVd) differ by five to six nucleotides located within the variable (V) domain referred to as the "cachexia expression motif". Sensitive hosts such as mandarin and its hybrids are seriously affected by cachexia disease. Current method...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2013.07.003
更新日期：2013-10-01 00:00:00
abstract：:DNA samples of 2303 individuals from nine different population groups were screened for variant -175g-->t in the promoter region of the low-density lipoprotein receptor (LDLR) gene. The -175g-->t variant detected at carrier frequencies of 3-10% in different African population groups was absent in the Caucasian and Asi...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/s0890-8508(03)00050-1
更新日期：2003-08-01 00:00:00
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
doi：10.1016/0890-8508(89)90004-2
更新日期：1989-09-01 00:00:00
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
doi：10.1016/j.mcp.2016.11.006
更新日期：2017-04-01 00:00:00
abstract：: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 d...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2005.05.003
更新日期：2005-10-01 00:00:00
