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 products in two stages. Ethidium bromide banding patterns characteristic of each of four subtypes of BKV were visualized through gel electrophoresis. A total of 37 samples from clinical specimens and culture fluids were successfully subtyped using the PCR-RE. A second method, PCR-sequencing, was applied to samples that generated less than 100 ng of DNA. These were subjected to a second round of PCR and then sequenced from single stranded templates immobilised via a biotinylated primer. The subtypes were assigned on the basis of the type-specific sequences previously characterized.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Jin Ldoi
10.1006/mcpr.1993.1047subject
Has Abstractpub_date
1993-08-01 00:00:00pages
331-4issue
4eissn
0890-8508issn
1096-1194pii
S0890-8508(83)71047-9journal_volume
7pub_type
杂志文章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::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
doi:10.1006/mcpr.1998.0166
更新日期:1998-06-01 00:00:00
abstract::Four isolates of Sarcocystis neurona from horses with equine protozoal myeloencephalitis and eight species of coccidia from the genera Sarcocystis, Toxoplasma or Eimeria were differentiated using the random amplified polymorphic DNA assay. A single, common, 550-bp DNA fragment was amplified from the DNA of each S. neu...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1994.1051
更新日期:1994-10-01 00:00:00
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
doi:10.1006/mcpr.2001.0384
更新日期:2001-12-01 00:00:00
abstract::The present study evaluates the usefulness of a PCR-based method for routine paternity testing in 35 paternity cases. This identification method which is based on amplification of three hypervariable genetic loci, apoB, D1S80 and HLA-DQ alpha, is compared, with regard to reliability and technical feasibility, to the c...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/0890-8508(92)90067-8
更新日期:1992-02-01 00:00:00
abstract::Staphylococcus aureus is responsible for a large panel of infections in humans and animals. In cows, S. aureus provokes chronic intramammary infections. Little information is available about the regulation of virulence factors in bovine isolates. Moreover, oxygenation, which is low in an inflamed mammary gland, could ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2005.01.002
更新日期:2005-08-01 00:00:00
abstract::alpha-SNAP is an essential component of the protein machinery responsible for membrane fusion events in different cell types. The hyh (hydrocephalus with hop gait) mouse carries a missense mutation in Napa gene that results in a point mutation (M105I) in alpha-SNAP protein. Homozygous animals for the mutant allele hav...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2009.07.002
更新日期:2009-12-01 00:00:00
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
doi:10.1006/mcpr.2002.0427
更新日期:2002-08-01 00:00:00
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
doi:10.1016/0890-8508(90)90013-p
更新日期:1990-04-01 00:00:00
abstract::Quantum dot (QD) molecular beacons were explored for sequence-specific DNA detection. The effectiveness of multiple linkage strategies and fluorescence quenchers were compared in hybridization-based assays. To compare linkage strategies, covalent amide linkage and streptavidin-biotin binding were used to link semicond...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2006.09.001
更新日期:2007-04-01 00:00:00
abstract::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...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/s0890-8508(06)80011-3
更新日期:1991-10-01 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::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
doi:10.1016/j.mcp.2012.11.001
更新日期:2013-04-01 00:00:00
abstract::Detection of Ehrlichia chaffeensis is necessary to study interactions between the parasite and its vertebrate and invertebrate hosts. The purpose of this study was to develop a sensitive, specific PCR assay for E. chaffeensis based on the outer membrane protein gene, p28. Candidate primer sets were identified and rank...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2003.10.002
更新日期:2004-04-01 00:00:00
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
doi:10.1016/j.mcp.2020.101541
更新日期:2020-06-01 00:00:00
abstract:BACKGROUND:Androgen receptor (AR) and long non-coding RNAs (lncRNA) play important roles in the initiation and progression of prostate cancer (PCa). The present study was designed to investigate whether lncRNA growth arrest-specific 5 (GAS5) is involved in the regulation of dexamethasone on the proliferation of AR+ PCa...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101607
更新日期:2020-10-01 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::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
doi:10.1006/mcpr.1994.1061
更新日期:1994-10-01 00:00:00
abstract::The ability to preserve and transport human cells in a stable medium over long distances is critical to collaborative efforts and the advancement of knowledge in the study of human disease. This is particularly important in the study of rare diseases. Recently, advancements in the understanding of renal ciliopathies h...
journal_title:Molecular and cellular probes
pub_type: 杂志文章,评审
doi:10.1016/j.mcp.2021.101694
更新日期:2021-01-08 00:00:00
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
doi:10.1016/j.mcp.2010.07.002
更新日期:2010-12-01 00:00:00
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
doi:10.1016/0890-8508(89)90015-7
更新日期:1989-12-01 00:00:00
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
doi:10.1016/j.mcp.2009.10.001
更新日期:2010-04-01 00:00:00
abstract::Homogenous fluorescence PCR assays offer distinct advantages for qualitative testing and are gaining immense popularity in fields like diagnostic microbiology. To meet the demand of high-volume laboratories, we developed a protocol for qualitative multiplex 5' nuclease assays using post-only PCR analysis. This novel a...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2005.06.001
更新日期:2005-10-01 00:00:00
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
doi:10.1016/j.mcp.2011.03.005
更新日期:2011-04-01 00:00:00
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
doi:10.1016/j.mcp.2006.03.007
更新日期:2006-12-01 00:00:00
abstract:OBJECTIVES:As one of the most serious malignant carcinomas that threaten the life of sufferers constantly, gastric cancer has attracted a lot of interest among researchers. miR-34a, a member of hundreds of microRNAs (miRNAs), has been elucidated to exert a suppressive role in gastric cancer tumorgenesis based on previo...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101601
更新日期:2020-10-01 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
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 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 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
doi:10.1016/j.mcp.2003.11.006
更新日期:2004-06-01 00:00:00