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 (amplicons). The u.v. and DNase treatments proved to be unsuitable for decontamination of PCR mixtures contaminated with amplicons shorter than 380 bp. By constructing special tube-holders and openers, and by applying a simple technique of pipetting, the false-positive PCR results were eliminated. The PCR products were visualized by three simple methods. The solid phase colorimetric method termed 'Detect Immobilized Amplified DNA' (DIANA) has been adapted to microplate. The other method, termed 'Colorimetric Detection Assay on Filter' (CODAF), proved to be very rapid. However, despite these advantages of DIANA and CODAF, henceforward the nucleic acid hybridization methods were found most reliable for safe identification of PCR amplicons. In order to simplify the hybridization, various non-radioactive labelling methods of oligonucleotide probes were compared. Biotinylation at the 5' end by means of oligonucleotide synthesis was the most simple and practical labelling method in this laboratory. The routine applicability of hybridization was further simplified by constructing a robot device, which automatically performs filter-hybridization and subsequently develops the signals derived from the biotinylated hybrids.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Belák S,Ballagi-Pordány Adoi
10.1006/mcpr.1993.1035subject
Has Abstractpub_date
1993-06-01 00:00:00pages
241-8issue
3eissn
0890-8508issn
1096-1194pii
S0890-8508(83)71035-2journal_volume
7pub_type
杂志文章,评审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
doi:10.1016/j.mcp.2006.10.007
更新日期:2007-06-01 00:00:00
abstract::The recently developed Inter-Simple Sequence Repeat PCR (ISSR-PCR) or microsatellite primed PCR or Simple Sequence Repeat (SSR)-Anchored PCR technique detects polymorphic markers in a wide variety of genomes. Usually the ISSR primers are either 5' end-labeled with gamma[32P]ATP or one of the alpha[32P] labeled dNTPs i...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.2001.0404
更新日期:2002-02-01 00:00:00
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
doi:10.1016/j.mcp.2005.05.002
更新日期:2005-10-01 00:00:00
abstract::The spectrum of cystic fibrosis (CF) mutations has been determined in many populations of different ethnic and geographic origins. However, in the south of Europe, the commonest mutation, delta F508, accounts for only about 50% of CF chromosomes, while identification of most of the other mutant alleles has not been ac...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/s0890-8508(95)80038-7
更新日期:1995-04-01 00:00:00
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
doi:10.1016/j.mcp.2004.03.001
更新日期:2004-08-01 00:00:00
abstract::The polymerase spiral reaction (PSR), a novel isothermal method for targeted DNA amplification, was effectively applied to detect Salmonella in artificially spiked pork. The specificity of the developed PSR was tested using 16 Salmonella and 15 non-Salmonella strains. The PSR assay was 10-fold more sensitive than conv...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101510
更新日期:2020-04-01 00:00:00
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
doi:10.1006/mcpr.1993.1017
更新日期:1993-04-01 00:00:00
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
doi:10.1016/j.mcp.2016.02.001
更新日期:2016-04-01 00:00:00
abstract::The identification and differentiation of the two variants of the ail gene, ailA from the more virulent American serotypes (08, 013a, 13b, 018, 020, 021) and ailNA from the less virulent non-American serotypes (03, 04, 05, 06, 09, 027 and 07, 8) was studied in a panel of 32 Yersinia enterocolitica human pathogenic iso...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1994.1025
更新日期:1994-06-01 00:00:00
abstract::Jersey haplotype (JH) 1, a stop-gain lethal mutation in the CWC15 gene, causes embryonic losses in Jersey cattle. Two PCR based assays using Amplification Refractory Mutation System (T-ARMS-PCR) and restriction fragment length polymorphism (PCR-RFLP) were developed for screening of the JH1 in cattle. During the screen...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101688
更新日期:2020-12-03 00:00:00
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
doi:10.1016/j.mcp.2016.07.004
更新日期:2016-10-01 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::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
doi:10.1016/j.mcp.2020.101636
更新日期:2020-10-01 00:00:00
abstract::This report describes detailed taxonomic and phylogenetic analysis of 15 non-tuberculous mycobacteria (NTMs) isolated from human pathological specimens in a Caribbean setting (12 slow-growers and three rapid-growers) that were not identified by cultural and biochemical tests and drug-susceptibility results. These isol...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2004.06.006
更新日期:2004-12-01 00:00:00
abstract::A rapid and sensitive recombinase polymerase amplification (RPA) assay, Bruce-RPA, was developed for detection of Brucella. The assay could detect as few as 3 copies of Brucella per reaction within 20 min. Bruce-RPA represents a candidate point-of-care diagnosis assay for human brucellosis. ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2016.02.007
更新日期:2016-04-01 00:00:00
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
doi:10.1006/mcpr.1997.0135
更新日期:1997-12-01 00:00:00
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
doi:10.1016/j.mcp.2013.12.003
更新日期:2014-08-01 00:00:00
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
doi:10.1006/mcpr.2000.0290
更新日期:2000-04-01 00:00:00
abstract::The development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for oth...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2019.02.002
更新日期:2019-04-01 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::Beak atrophy and dwarfism syndrome (BADS) is commonly caused by co-infection with duck circovirus (DuCV) and novel goose parvovirus (NGPV). Therefore, concurrent detection of both viruses is important for monitoring and limiting BADS, although such a diagnostic test has not been reported. In this study, we developed a...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101648
更新日期:2020-10-01 00:00:00
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
doi:10.1016/j.mcp.2007.06.010
更新日期:2008-02-01 00:00:00
abstract::Three species of Dermacentor, Dermacentor albipictus, Dermacentor andersoni and Dermacentor variabilis, commonly occur in Canada. D. andersoni and D. variabilis are morphologically similar and are important vectors of human and animal pathogens. A practical polymerase chain reaction (PCR) assay, based on the amplifica...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2007.04.003
更新日期:2007-10-01 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::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
doi:10.1016/j.mcp.2015.08.007
更新日期:2015-12-01 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::The molecular tag vaccine against Brucella abortus and serological testing are the main methods of prevention of brucellosis used currently. They can discriminate vaccinated animals and humans from those naturally infected. In this study, we constructed a gene deletion mutant strain, B. abortus S19 virB5 with a molecu...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2012.02.003
更新日期:2012-06-01 00:00:00
abstract::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 dideoxynucleoti...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1995.0027
更新日期:1995-06-01 00:00:00
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
doi:10.1016/j.mcp.2004.07.006
更新日期:2005-02-01 00:00:00
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
doi:10.1016/j.mcp.2017.06.005
更新日期:2017-10-01 00:00:00