Abstract：
:Citrus plants are natural hosts of five viroid species and a large number of sequence variants. Because of their small size, viroids lend themselves to various RT-PCR approaches for their detection and further characterization. The one-step RT-PCR approach proposed here is based on the synthesis of viroid-cDNA by reverse transcription at 60 degrees C using a viroid specific 27-mer primer followed by standard second strand synthesis plus PCR amplification with various primer pairs. According to the primers used, full or partial length viroid-DNA is obtained. The technique avoids amplicon contamination in routine diagnosis. The suitability of the technique has been demonstrated using several nucleic acid extraction procedures and different viroid infected host species. The homogenization of tissue inside sealed plastic bags followed by nucleic acid extraction using a SDS/potassium acetate method is recommended because of its efficiency, simplicity and low cost. This extraction procedure, when coupled to the one-step RT-PCR approach, can be useful to avoid cross-contamination during routine diagnosis. A PCR strategy capable of discriminating between mild and severe strains of CEVd and identifying cachexia-inducing isolates of HSVd, is also described.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Bernad L,Duran-Vila Ndoi
10.1016/j.mcp.2005.11.001subject
Has Abstractpub_date
2006-04-01 00:00:00pages
105-13issue
2eissn
0890-8508issn
1096-1194pii
S0890-8508(05)00096-4journal_volume
20pub_type
杂志文章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 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
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：: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：:A polymorphic GT dinucleotide repeat sequence has been identified in the 5' flanking region of the human growth hormone receptor (hGHR) gene on chromosome 5p13.1-p12, within the promoter region of the V9 5'UTR exon. Thirteen alleles have been identified in 50 non-related individuals, with an observed heterozygosity of...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.2001.0366
更新日期：2001-08-01 00:00:00
abstract：:In situ hybridization is the hybridization-mediated detection of specific nucleic acid sequences within structurally intact cells or tissues. As such it uniquely provides localization of nucleic acid superimposed on observable cellular and subcellular structural detail, allowing analysis unobtainable by other hybridiz...
journal_title：Molecular and cellular probes
pub_type： 杂志文章,评审
doi：10.1016/0890-8508(87)90033-8
更新日期：1987-09-01 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：: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
doi：10.1016/0890-8508(90)90023-s
更新日期：1990-08-01 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：: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：:To produce hybrids, one member of the parental line is genetically made male-sterile. This male-sterile trait is encoded by mitochondria so that it is maternally inherited. Consequently, the progeny of a male-sterile plant is fully sterile. Nevertheless, during the handling of cytoplasmic male-sterile seed stocks, som...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(91)90032-f
更新日期：1991-02-01 00:00:00
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
doi：10.1016/j.mcp.2016.01.003
更新日期：2016-02-01 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：: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：:The present report describes a real-time PCR-based procedure to reliably determine the quantity of Leishmania amastigotes in relation to the amount of host tissue in histological skin sections from canine and equine cases of cutaneous leishmaniasis. The novel diagnostic Leishmania-PCR has a detection limit of <0.02 am...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2015.09.008
更新日期：2015-12-01 00:00:00
abstract：:We have developed a highly specific and sensitive technique for the detection of Pneumocystis carinii DNA using DNA amplification by the polymerase chain reaction (PCR). PCR products are detected by agarose gel electrophoresis and Southern hybridization to an oligonucleotide probe. Here we report the calibration of pa...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(92)90055-3
更新日期：1992-04-01 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：: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：:The aim of the present study was to evaluate a mixed phase DNA hybridization assay for detection of Bordetella pertussis and Bordetella parapertussis in nasopharyngeal aspirates from patients with suspected pertussis. Among 179 consecutive patients with own or parental suspicion of pertussis, the diagnosis was confirm...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(90)90021-q
更新日期：1990-08-01 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：:Variable number tandem repeat (VNTR) is a frequently employed typing method of Mycobacterium avium paratuberculosis (MAP) isolates. Based on whole genome sequencing in a previous study, allelic diversity at some VNTR loci seems to over- or under-estimate the actual phylogenetic variance among isolates. Interestingly, ...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2016.02.002
更新日期：2016-06-01 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：: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
doi：10.1016/j.mcp.2017.11.002
更新日期：2018-02-01 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：:Pre-mRNA processing factor 4 (PRPF4), a core protein in U4/U6 snRNP, maintains snRNP structures by interacting with PRPF3 and cyclophilin H. Expression of the PRPF4 gene affects cell survival as well as apoptosis and is responsible for retinitis pigmentosa (RP). Proteomics analysis shows that PRPF4 may be a therapeuti...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2019.101440
更新日期：2019-10-01 00:00:00
abstract：:The phenotypic detection of Campylobacter concisus, a species of considerable genomic and phenotypic heterogeneity, has proven to be rather tedious in the past. Although alternative methods like DNA:DNA hybridization, immunotyping or whole-cell protein electrophoresis are valuable for the specific detection of C. conc...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/s0890-8508(95)90114-0
更新日期：1995-08-01 00:00:00
abstract：:The flagellin genes of four Burkholderia pseudomallei and two Burkholderia cepacia clinical isolates were studied by a polymerase chain reaction (PCR)-based isolation method using the same pair of primers. The PCR-amplification products of the isolates showed a single band of about 1.1 kb, which is similar to a type I...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1999.0221
更新日期：1999-04-01 00:00:00
abstract：:Members of the Mycobacterium chelonae complex (MCC), namely M. chelonae, Mycobacterium abscessus and Mycobacterium immunogenum, have been implicated in nosocomial infections and occupational respiratory illnesses like hypersensitivity pneumonitis (HP) associated with contaminated metalworking fluid (MWF) exposures. Cl...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2004.09.007
更新日期：2005-04-01 00:00:00
abstract：:To find a specific method for HLA-B27 typing, we tested an HLA-B27-specific polymerase chain reaction. This method was used for screening 100 randomly selected blood donors, 10 of them being HLA-B27 positive. A flow cytometric method and this PCR method were compared. ...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1997.0112
更新日期：1997-08-01 00:00:00
abstract：:TTR amyloidosis (ATTR) is a fatal condition caused by extracellular deposits of misfolded transthyretin. Patients often present with cardiac disease, but manifestations may also involve other organs including the peripheral nervous system. ATTR is considered familial when heterozygous mutations in the TTR gene are pre...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2018.08.005
更新日期：2018-10-01 00:00:00
