Abstract:
:A duplex real-time SYBR Green LightCycler PCR (LC-PCR) assay with DNA extraction using the QIAamp DNA Stool Mini kit was evaluated with regard to detection of 8 of 17 species of food- or waterborne pathogens in five stool specimens in 2 h or less. The protocol used the same LC-PCR with 20 pairs of specific primers. The products formed were identified based on a melting point temperature (T(m)) curve analysis. The 17 species of food- or waterborne pathogens examined were enteroinvasive Escherichia coli, enteropathogenic E. coli, enterohemorrhagic E. coli, enterotoxigenic E. coli, enteroaggregative E. coli, Salmonella spp., Shigella spp., Yersinia enterocolitica, Yersinia pseudotuberculosis, Campylobacter jejuni, Vibrio cholerae, Vibrio parahaemolyticus, Vibrio vulnificus, Aeromonas spp., Staphylococcus aureus, Clostridium perfringens, and Bacillus cereus. No interference with the LC-PCR assay was observed when stool specimens were artificially inoculated with each bacterial species. The detection levels were approximately 10(5) food- or waterborne pathogenic bacteria per g of stool. The protocol for processing stool specimens for less than 10(4) food- or waterborne pathogenic bacteria per g of stool requires an overnight enrichment step to achieve adequate sensitivity. However, the rapid amplification and reliable detection of specific genes of greater than 10(5) food- or waterborne pathogenic bacteria per g in samples should facilitate the diagnosis and management of food- or waterborne outbreaks.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Fukushima H,Tsunomori Y,Seki Rdoi
10.1128/jcm.41.11.5134-5146.2003subject
Has Abstractpub_date
2003-11-01 00:00:00pages
5134-46issue
11eissn
0095-1137issn
1098-660Xjournal_volume
41pub_type
杂志文章abstract::PCR-restriction fragment length polymorphism analysis (PRA) of the hsp65 gene (A. Telenti, F. Marchesi, M. Balz, F. Bally, E. C. Böttger, and T. Bodmer, J. Clin. Microbiol. 31:175-178, 1993) was applied to 108 mycobacterial isolates representing 34 species to evaluate its potential as a rapid reference method. A total...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.11.2969-2973.1997
更新日期:1997-11-01 00:00:00
abstract::We evaluated the performance of the DR. TBDR/NTM IVD kit, which was designed to detect Mycobacterium tuberculosis, rifampin-resistant M. tuberculosis, and nontuberculous mycobacteria, for detecting 110 positive and 50 negative cultures in Mycobacterium Growth Indicator Tubes. The accuracy rate of this kit for identifi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01862-12
更新日期:2012-10-01 00:00:00
abstract::Longitudinal data on the E6/E7 mRNA-based Aptima human papillomavirus (AHPV) assay exceeding three years in comparison to the gold standard Digene Hybrid Capture 2 (HC2) test are not available. We previously reported the cross-sectional data of the German AHPV Screening Trial (GAST) in which 10,040 women were recruite...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01177-18
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abstract::A 1-year prospective study was carried out in two large urban centers of São Paulo State, Brazil, to determine the prevalences and roles of the different Escherichia coli pathotypes in children less than 5 years of age with diarrhea presenting to the emergency rooms of public hospitals or visiting private pediatrician...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00084-07
更新日期:2007-10-01 00:00:00
abstract::For a total of 29 non-human immunodeficiency virus 1 cryptococcal meningitis cases, titer changes in the latex agglutination test before and after therapy were reviewed along with clinical manifestations, laboratory findings, and therapy regimens. The cryptococcal antigen titer decreased for every case after therapy a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.6.2989-2990.2005
更新日期:2005-06-01 00:00:00
abstract::Although rotavirus genotype P[6] is one of the three most common VP4 specificities associated with human infection, the relatively few sequence data available in public databases suggest that the genetic variability within P[6] might be presently unexplored. Thus far, two human P[6] lineages (M37-like and AU19-like) a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.9.4338-4343.2004
更新日期:2004-09-01 00:00:00
abstract::In order to better understand the role of Mycobacterium leprae nasal carriage in the maintenance of infection reservoirs and transmission of leprosy, we applied a polymerase chain reaction (PCR) that detected a 531-bp fragment of the pra gene of M. leprae on nasal swab specimens collected through a total population su...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.11.2947-2951.1993
更新日期:1993-11-01 00:00:00
abstract::Developing interpretive breakpoints for any given organism-drug combination requires integration of the MIC distribution, pharmacokinetic and pharmacodynamic parameters, and the relationship between the in vitro activity and outcome from both in vivo and clinical studies. Using data generated by standardized broth mic...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.44.3.819-826.2006
更新日期:2006-03-01 00:00:00
abstract::Murine monoclonal antibodies (MAbs) against herpes simplex virus type 1 and 2 (HSV-1 and -2, respectively) nuclear antigens were used to identify cells infected with HSV-1 or -2 by indirect immunofluorescence in conjunction with flow cytometry after virus amplification of MRC-5 cell monolayers. The results indicate th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.8.1864-1866.1990
更新日期:1990-08-01 00:00:00
abstract::Four slightly yellow-pigmented, alpha-hemolytic, gram-negative coccobacilli, three from wound specimens and one from multiple blood cultures of a patient with endocarditis, were identified as Neisseria elongata subsp. glycolytica on the basis of their overall biochemical and genetic similarities to this subspecies. Th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.1.76-78.1995
更新日期:1995-01-01 00:00:00
abstract::We have developed a method for determining the serotypes of poliovirus isolates by PCR. Three sets of serotype-specific antisense PCR-initiating primers (primers seroPV1A, seroPV2A, and seroPV3A) were designed to pair with codons of VP1 amino acid sequences that are conserved within but that differ across serotypes. T...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.2.352-357.1998
更新日期:1998-02-01 00:00:00
abstract::Neisseria meningitidis is the causative agent of meningococcal sepsis and meningitis. Neisseria polysaccharea is a nonpathogenic species. N. polysaccharea is able to use sucrose to produce amylopectin, a starch-like polysaccharide, which distinguishes it biochemically from the pathogenic species N. meningitidis. The d...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.1.273-278.2003
更新日期:2003-01-01 00:00:00
abstract::The serum antibody response of infants and children immunized with Formalin-inactivated respiratory syncytial virus (RSV) vaccine 20 years ago was determined by using an enzyme-linked immunosorbent assay specific for the RSV fusion (F) and large (G) glycoproteins and a neutralization assay. Twenty-one young infants (2...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.24.2.197-202.1986
更新日期:1986-08-01 00:00:00
abstract::A standardized broth microdilution method was compared to the E test and an agar dilution method for the antimicrobial susceptibility testing of Campylobacter jejuni and C. coli isolates. A group of 47 human clinical isolates, 37 isolates from retail poultry, and 29 isolates from living turkeys (total, 113 isolates) w...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.3.1062-1068.2003
更新日期:2003-03-01 00:00:00
abstract::Of 522 patients with Propionibacterium acnes bacteremia (PAB), 18 (3.5%) had clinically significant PAB. Of these 18 patients, 10 (55.6%) had hospital-acquired bacteremia and 6 (33.3%) had undergone invasive procedures before development of PAB. One patient with a ventricular septal defect presented with infective end...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01842-10
更新日期:2011-04-01 00:00:00
abstract::Droplet digital PCR (ddPCR) is an emulsion PCR process that performs absolute quantitation of nucleic acids. We developed a ddPCR assay for Chlamydia trachomatis infections and found it to be accurate and precise. Using PCR mixtures containing plasmids engineered to include the PCR target sequences, we were able to qu...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00622-13
更新日期:2013-07-01 00:00:00
abstract::Taeniasis due to Taenia solium is a disease with important public health consequences, since the larval stage is not exclusive to the animal intermediate, the pig, but also infects humans, causing neurocysticercosis. Early diagnosis and treatment of T. solium tapeworm carriers is important to prevent human cysticercos...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01172-07
更新日期:2008-01-01 00:00:00
abstract::Freshly isolated clinical strains of Klebsiella were tested for the ability to fix dinitrogen by the acetylene reduction assay. Ability to detect this trait was markedly affected by cultural conditions. When the test was run at 37 degrees C in the presence of yeast extract (50 mg/liter), only 1.6% of the organisms wer...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1977-11-01 00:00:00
abstract::A 24-year-old man sustained a crush injury to the tibia, which subsequently became infected with Saksenaea vasiformis. He was treated with debridement and a free myocutaneous flap, but amputation was necessary because of mycotic osteomyelitis. S. vasiformis was recovered and identified on the basis of its characterist...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.5.933-935.1987
更新日期:1987-05-01 00:00:00
abstract::We sequenced homologous chromosomal loci from several North American and European isolates of the Lyme disease spirochete Borrelia burgdorferi, as well as from the relapsing fever spirochete Borrelia hermsii. Inter- and intraspecies sequence comparisons permitted the design of B. burgdorferi-specific polymerase chain ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.3.524-532.1991
更新日期:1991-03-01 00:00:00
abstract::The sensitivity and specificity of the human immunodeficiency virus (HIV) type 1-specific immunoglobulin G capture enzyme-linked immunosorbent assay (BED-CEIA) were compared with those of the avidity index method to identify recent HIV infection using a panel of 148 samples (81 patients) representing durations of infe...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01055-07
更新日期:2008-01-01 00:00:00
abstract::Although antiretroviral treatment availability has improved, the virological monitoring of patients remains largely uneven across regions. In addition, viral quantification tests are suffering from human immunodeficiency virus type 1 (HIV-1) genetic diversity, fueled by the emergence of new recombinants and of lentivi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02792-12
更新日期:2013-03-01 00:00:00
abstract::A modified CAMP test was used to identify 973 Vibrio cholerae isolates by phenotype. Eltor and non-O1 strains were CAMP positive; classical strains were CAMP negative. Sausage-shaped zones of hemolysis of eltor strains were easily distinguished from narrower bands of non-O1 isolates. For O1 isolates, there was 100% ag...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.32.1.235-237.1994
更新日期:1994-01-01 00:00:00
abstract::Nosocomial transmission of Neisseria meningitidis has only rarely been reported. Here, we present a significant spatiotemporal association of two cases of invasive meningococcal disease identified by retrospective cluster analysis with the program SaTScan. The most likely epidemiological link was simultaneous hospital...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00613-06
更新日期:2006-11-01 00:00:00
abstract::Leptospira is a highly diverse genus comprising many species and serogroups in Brazil as well as all over the world. However, a study by arbitrarily primed PCR of 44 leptospiral strains isolated from humans during three different outbreaks in Brazilian urban centers reveals that 43 of 44 isolates exhibit very similar ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:2000-01-01 00:00:00
abstract::Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid amplification method in which reagents react rapidly and efficiently, with a high specificity, under isothermal conditions. We used a LAMP assay for the detection of herpes simplex virus type 1 (HSV-1), herpes simplex virus type 2 (HSV-2), and varic...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.7.3290-3296.2005
更新日期:2005-07-01 00:00:00
abstract::A batch of fetal calf serum was encountered which suppressed the number of Chlamydia trachomatis inclusions seen when it was used in conjunction with cycloheximide-treated McCoy cells. The batch of serum did not affect the number of inclusions detected in irradiated or untreated McCoy cells. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.11.4.424-425.1980
更新日期:1980-04-01 00:00:00
abstract::Bacteroides forsythus has been associated with destructive adult periodontitis. Up to now, detailed analysis by classical means was hampered by the fastidious nature of the organism. There is hope that the application of molecular detection methods such as indirect immunofluorescence or in situ hybridization (ISH) wil...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.4.941-946.1993
更新日期:1993-04-01 00:00:00
abstract::Studies were conducted to compare and evaluate antimicrobial susceptibility test results for enterococci obtained by six national committee disk diffusion procedures. Variations in the incidence of isolates in resistance categories and errors were associated with the use of ciprofloxacin, gentamicin, nitrofurantoin, r...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.10.3753-3756.2001
更新日期:2001-10-01 00:00:00
abstract::Biotype -specificity of Staphylococcus aureus of human and canine origins has been found to be associated with thermolabile agglutinogens represented in S. aureus strains 17 and 61218, respectively. Both strains also have exhibited a common thermostable antigen. On that basis, absorbed antisera have been developed for...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.21.1.43-45.1985
更新日期:1985-01-01 00:00:00