Abstract:
:A rapid and highly sensitive real-time PCR detection and quantification method for infectious hypodermal and hematopoietic necrosis virus (IHHNV), a single-stranded DNA virus, and white spot virus (WSV), a double-stranded DNA (dsDNA) virus infecting penaeid shrimp (Penaeus sp.), was developed using the GeneAmp 5700 sequence detection system coupled with SYBR Green chemistry. The PCR mixture contains a fluorescence dye, SYBR Green, which upon binding to dsDNA exhibits fluorescence enhancement. The enhancement of fluorescence was proportional to the initial concentration of the template DNA. A linear relationship was observed between the amount of input plasmid DNA and cycle threshold (C(T)) values over a range of 1 to 10(5) copies of the viral genome. To control the variation in sampling and processing among samples, the shrimp beta-actin gene was amplified in parallel with the viral DNA. The C(T) values of IHHNV- and WSV-infected samples were used to determine absolute viral copy numbers from the standard C(T) curves of these viruses. For each virus and its beta-actin control, the specificity of amplification was monitored by using the dissociation curve of the amplified product. Using genomic DNA as a template, SYBR Green PCR was found to be 100- to 2000-fold more sensitive than conventional PCR, depending on the virus, for the samples tested. The results demonstrate that SYBR Green PCR can be used as a rapid and highly sensitive detection and quantification method for shrimp viruses and that it is amenable to high-throughout assay.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Dhar AK,Roux MM,Klimpel KRdoi
10.1128/JCM.39.8.2835-2845.2001subject
Has Abstractpub_date
2001-08-01 00:00:00pages
2835-45issue
8eissn
0095-1137issn
1098-660Xjournal_volume
39pub_type
杂志文章abstract::Species of the genus Bartonella are currently recognized in growing numbers and are involved in an increasing variety of human diseases, mainly trench fever, Carrion's disease, bacillary angiomatosis, endocarditis, cat scratch disease, neuroretinitis, and asymptomatic bacteremia. Such a wide spectrum of infections mak...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.8.2768-2778.2001
更新日期:2001-08-01 00:00:00
abstract::In a survey of bloodstream infection (BSI) isolates across the continental United States, 162 Candida albicans isolates were fingerprinted with the species-specific probe Ca3 and the patterns were analyzed for relatedness with a computer-assisted system. The results demonstrate that particular BSI strains are more hig...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.6.1518-1529.1998
更新日期:1998-06-01 00:00:00
abstract::Evaluation of previously described chemically defined media for the growth of Legionella pneumophila showed that these media supported poor growth of several strains of L. pneumophila and did not support growth of certain of the Legionella species described later. Growth was stimulated by the dialysate from yeast extr...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.1.163-169.1986
更新日期:1986-01-01 00:00:00
abstract::tRNA intergenic spacer PCR (tDNA-PCR) was evaluated for its usefulness in the differentiation of enterococcal species of human and animal origin. This technique was carried out for 124 strains belonging to 17 enterococcal species and generated DNA fragments, which were separated by capillary electrophoresis. tDNA-PCR ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.11.4201-4207.2000
更新日期:2000-11-01 00:00:00
abstract::All but 2 of 63 Mycobacterium avium isolates from distinct geographic areas of Italy exhibited markedly polymorphic, multibanded IS1245 restriction fragment length polymorphism (RFLP) patterns; 2 isolates showed the low-number banding pattern typical of bird isolates. By computer analysis, 41 distinct IS1245 patterns ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.12.3694-3697.1998
更新日期:1998-12-01 00:00:00
abstract::The aim of this study was to evaluate the seroprevalence of human herpesvirus 8 (HHV-8) in a group of 150 patients awaiting heart transplants and to detect HHV-8 seroconversion after transplantation. Four patients were HHV-8 seropositive before transplantation, and one of them developed Kaposi's sarcoma. One patient c...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.6.2217-2219.2002
更新日期:2002-06-01 00:00:00
abstract::Helicobacter pylori is known to be a major cause of peptic ulceration. The jhp0562 gene, encoding a glycosyltransferase involved in the synthesis of the lipopolysaccharide, was associated with peptic ulcer disease (PUD) in children. The beta-(1,3)-galactosyltransferase [beta-(1,3)GalT] gene (jhp0563), involved in Lewi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00401-10
更新日期:2010-08-01 00:00:00
abstract::Two national surveys were conducted to describe the incidence and prevalence of Enterobacter aerogenes in 21 Belgian hospitals in 1996 and 1997 and to characterize the genotypic diversity and the antimicrobial resistance profiles of clinical strains of E. aerogenes isolated from hospitalized patients in Belgium in 199...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.3.889-896.2001
更新日期:2001-03-01 00:00:00
abstract::A pseudooutbreak of cryptococcal meningitis was caused by the use of contaminated albumin solution used in the preparation of Papanicolaou-stained slides of cerebrospinal fluid. Organisms were seen in cytocentrifuge preparations, but not in India ink preparations of cerebrospinal fluid specimens. Cryptococcal antigen ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.15.6.1141-1143.1982
更新日期:1982-06-01 00:00:00
abstract::Molecular types of the Cryptococcus neoformans/Cryptococcus gattii species complex that infect dogs and cats differ regionally and with host species. Antifungal drug susceptibility can vary with molecular type, but the susceptibility of Cryptococcus isolates from dogs and cats is largely unknown. Cryptococcus isolates...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03392-13
更新日期:2014-06-01 00:00:00
abstract::A procedure for detecting pneumococcal capsular polysaccharides in immune complexes is described. Separation of antigen from immune complexes is achieved by electrophoresis at 56 degrees C. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1978-08-01 00:00:00
abstract::Influenza viruses related to the markedly antigenically divergent strains B/Yamagata/16/88 and B/Victoria/2/87 are circulating in human populations. Adults develop cross-reacting antibodies against recent and earlier influenza B virus strains after vaccination with B/Yamagata/16/88, probably because of previous influe...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.7.1530-1532.1991
更新日期:1991-07-01 00:00:00
abstract::Whole-cell chromosomal digests of 54 isolates of Streptococcus suis encompassing all known serotypes from a geographically varied collection were examined by PstI restriction fragment length polymorphisms and then hybridized with a digoxigenin-11-dUTP-labeled cDNA probe transcribed from a mixture of 16S and 23S rRNAs ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.4.968-972.1995
更新日期:1995-04-01 00:00:00
abstract::Two in a group of five naturally seropositive donkey stallions were found to shed equine arteritis virus (EAV) in their semen as demonstrated by virus isolation. Direct intramuscular inoculation of sonicated semen from one virus-shedding stallion (S3) caused clinical disease in two donkeys from which virus was recover...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.12.3296-3299.1995
更新日期:1995-12-01 00:00:00
abstract::Of 12,321 stool samples analyzed over a 6-year interval, 870 (7.1%) were positive for a total of 1,019 parasites, of which 1,011 (99.2%) were found in trichrome-stained smears of unconcentrated specimens while only 479 (47.0%) were detected in iodine-stained smears of concentrated samples. Stool specimens were next an...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.3.835-837.1999
更新日期:1999-03-01 00:00:00
abstract::A rapid method for detection of respiratory syncytial virus (RSV) in nasopharyngeal aspirates, involving a combination of reverse transcription and polymerase chain reaction amplification (RT-PCR), has been developed. The RT-PCR assay employs oligonucleotide primers specific for the region of the RSV genome which enco...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.4.901-904.1992
更新日期:1992-04-01 00:00:00
abstract::An apparent outbreak of serious Candida albicans infections (n = 6) occurred in a surgical intensive care unit over a 4-week period. Four patients developed C. albicans bloodstream infections. An additional patient developed catheter-related C. albicans infection; the sixth patient developed an infection of cerebrospi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.3.576-580.1995
更新日期:1995-03-01 00:00:00
abstract::The efficacy of antiviral treatment for chronic hepatitis C virus (HCV) infection is determined by measuring HCV RNA at specific time points throughout therapy using highly sensitive and accurate HCV RNA assays. This study compared the performances of two recently developed real-time PCR HCV RNA assays, cobas HCV for ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.02193-16
更新日期:2017-04-01 00:00:00
abstract::Nineteen Rhizomucor miehei and Rhizomucor pusillus isolates were assayed for their ability to utilize 87 various substrates as a single carbon source. Besides a difference in sucrose utilization, distinctive differences were found in the utilization of glycine, phenylalanine, and beta-alanine. Five isoenzyme systems a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.7.2153-2156.1998
更新日期:1998-07-01 00:00:00
abstract::Four commercial kits, Oxi/Ferm (OF), API 20E (AP), Minitek (MT; BBL Microbiology Systems), and Flow N/F (NF), were evaluated, without additional tests, for identification of 258 gram-negative nonfermentative rods. OF and MT were read after 48 h of incubation, and AP and NF were read after both 24 and 48 h of incubatio...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.12.2.271-278.1980
更新日期:1980-08-01 00:00:00
abstract::We determined the entire nucleotide sequence of phiSa2958-carrying Panton-Valentine leukocidin (PVL) gene, which was lysogenized in a sequence type 5 staphylococcal cassette chromosome mec (SCCmec) type II strain of methicillin-resistant Staphylococcus aureus (MRSA). Based on the nucleotide sequences of PVL phages, we...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00136-08
更新日期:2008-10-01 00:00:00
abstract::A DNA microarray platform, based on the nucleotide sequences of the internal transcribed spacer regions (ITS1 and ITS2) of the rRNA gene, was developed to identify 32 fungal pathogens at the species level. The probe sequences were spotted onto polycarbonate slides with a mini-microarray printer, and after the hybridiz...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01908-17
更新日期:2018-04-25 00:00:00
abstract::The Tigecycline Evaluation and Surveillance Trial (TEST) was designed to monitor global longitudinal changes in bacterial susceptibility to a panel of antimicrobial agents, including tigecycline. In this study, we examine susceptibility among Gram-positive isolates collected from pediatric patients globally between 20...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00157-13
更新日期:2013-07-01 00:00:00
abstract::Marburg virus (MARV) causes a severe hemorrhagic fever in humans with a high mortality rate. The rapid and accurate identification of the virus is required to appropriately provide infection control and outbreak management. Here, we developed and evaluated a one-step reverse transcription-loop-mediated isothermal ampl...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01224-09
更新日期:2010-07-01 00:00:00
abstract::The food-borne pathogen Listeria monocytogenes is genetically heterogeneous. Although some clonal groups have been implicated in multiple outbreaks, there is currently no consensus on how "epidemic clones" should be defined. The objectives of this work were to compare the patterns of sequence diversity on two sets of ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01874-13
更新日期:2013-11-01 00:00:00
abstract::The usefulness of filter paper for preservation of bacterial cells was shown by mixed-linker DNA fingerprint analysis of Mycobacterium tuberculosis isolates from 77 Brazilian patients. DNA fingerprints of samples spotted onto filter paper and conventional culture material were identical. Thus, filter paper specimens a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.2.573-576.1998
更新日期:1998-02-01 00:00:00
abstract::We compared the microhemagglutination assay for Treponema pallidum (MHA-TP), a treponemal test, with two other treponemal tests, the Serodia Treponema pallidum particle agglutination (TP-PA) assay and the Captia Syphilis-G enzyme immunoassay, using 390 clinical serum samples. We also compared two nontreponemal tests, ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.7.2543-2545.2000
更新日期:2000-07-01 00:00:00
abstract::In clinical laboratories, diagnosis of imported malaria is commonly performed by microscopy. However, the volume of specimens is generally low and maintaining proficiency in reading blood smears, particularly at the species level, is challenging in this setting. To address this problem, the Provincial Laboratory for P...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00195-13
更新日期:2013-06-01 00:00:00
abstract::A total of 28 unrelated isolates of the Salmonella enterica subsp. enterica serovar dublin (S. dublin) collected during a 6-year period, as well as four samples of the S. dublin live vaccine strain Bovisaloral and its prototype strain S. dublin 442/039, were investigated by different molecular typing methods for the f...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.3.641-646.1996
更新日期:1996-03-01 00:00:00
abstract::Serratia ficaria was first described in 1979 as part of the fig tree ecosystem (P.A.D. Grimont, F. Grimont, and M. P. Starr, Curr. Microbiol. 2:277-282, 1979). Since then, it has been isolated from clinical specimens from a few human patients (C. Bollet, J. Freney, P. de Micco, F. Grimont, and P.A.D. Grimont, Méd. Mal...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
doi:10.1128/JCM.32.9.2285-2288.1994
更新日期:1994-09-01 00:00:00