Abstract:
:PCR-based molecular assays have a central role in polyomavirus diagnostics. To assure optimal performance, target sequences should be regularly updated according to newly available sequences. The aim of this study was to review our in-house polyomavirus BK (BKV) and JC (JCV) real-time PCR assays. Database analysis revealed variations in the BKV target region which might affect the assay performance, while no significant changes were found in the JCV target region. We compared two degenerate versions of our BKV primers which accommodated at least 95% of all published genetic variants. Dilutions of cloned viral genomic DNA and probit analysis indicated an analytical sensitivity of the updated BKV assay of 4.15 copies/reaction and that of the JCV assay was 3.37 copies/reaction. The specificity was assessed by testing JCV- and BKV-positive samples that showed no cross-reactivity. The performance of the original and updated BKV assay was compared in 101 urine and 200 plasma samples submitted to our routine diagnostic laboratory revealed similar quantitative results. We conclude that our JCV and updated BKV real-time PCR assays are robust and detect rare variants possibly encountered in the clinical routine.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Dumoulin A,Hirsch HHdoi
10.1128/JCM.02008-10subject
Has Abstractpub_date
2011-04-01 00:00:00pages
1382-8issue
4eissn
0095-1137issn
1098-660Xpii
JCM.02008-10journal_volume
49pub_type
杂志文章abstract::Baculovirus-expressed foot-and-mouth disease virus (FMDV) nonstructural protein 3AB was used as the antigen in an enzyme-linked immunosorbent assay. This assay allowed the differentiation of vaccinated from infected pigs. Serial studies were performed using sera collected from pigs in the field. Positive reactions wer...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.8.2843-2848.2002
更新日期:2002-08-01 00:00:00
abstract::Methicillin-resistant Staphylococcus aureus (MRSA) infection and colonization, involving MRSA strains which differ from common human health care-associated clones, have become serious emerging conditions in equine veterinary hospitals. In 2010, MRSA spa type t535 caused an outbreak involving both horses and personnel ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00090-15
更新日期:2015-09-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02573-06
更新日期:2007-09-01 00:00:00
abstract::We report a failure of the real-time reverse transcriptase PCR H7 subtyping protocol currently used in national avian influenza surveillance programs. Significant substitutions in primer and probe target sequences were identified, especially in wild bird viruses. The protocol, originally designed for detecting H7 infl...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02426-07
更新日期:2008-05-01 00:00:00
abstract::Parasite-specific humoral and cellular immune responses were evaluated in in nine immunosuppressed patients with Strongyloides stercoralis infection. Four patients had the disseminated form of the disease, and five had serious manifestations without evidence of hyperinfection. All patients had high levels of immunoglo...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.6.1099-1103.1986
更新日期:1986-06-01 00:00:00
abstract::A public health response relies upon rapid and reliable confirmation of disease by diagnostic assays. Here, we detail the design and validation of two variola virus-specific real-time PCR assays, since previous assays cross-reacted with newly identified cowpox viruses. The assay specificity must continually be reasses...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.03613-14
更新日期:2015-04-01 00:00:00
abstract::The reliability of the novel BACTEC MGIT 960 pyrazinamide (PZA) kit (Becton Dickinson Microbiology Systems, Sparks, Md.) was assessed for testing of susceptibility of Mycobacterium tuberculosis to PZA. Results generated by the BACTEC MGIT 960 system (Becton Dickinson) were compared with those obtained with the BACTEC ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.5.1670-1674.2002
更新日期:2002-05-01 00:00:00
abstract::Total DNA from five Campylobacter species was digested with a mixture of XhoI and BglII restriction endonucleases and analyzed by Southern hybridization by using a probe complementary to the DNA coding for the 16S rRNA. Each of the Campylobacter species, including C. jejuni, C. coli, C. laridis, C. fetus, and C. upsal...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.7.1514-1517.1989
更新日期:1989-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.6.1194-1197.1988
更新日期:1988-06-01 00:00:00
abstract::The detection and identification of intestinal helminths were studied retrospectively by comparing the Formalin concentration technique with the trichrome-stained smear technique. A total of 3,997 stool samples from 1,570 patients were examined by both methods. Of the 3,997 samples, 31% (1,239 of 3,997) contained helm...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.16.6.1137-1144.1982
更新日期:1982-12-01 00:00:00
abstract::A convenient assay combining solution hybridization and enzyme immunoassay for DNA-RNA hybrids (polymerase chain reaction-enzyme immunoassay [PCR-EIA]) was developed to detect human immunodeficiency virus type 1 (HIV-1) provirus amplified by the PCR and was compared with oligomer hybridization with 32P-labeled SK19. I...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.11.2461-2467.1991
更新日期:1991-11-01 00:00:00
abstract::Immunoblots were used to study the immunoglobulin G response to Borrelia burgdorferi in experimentally and naturally exposed dogs. Adsorption studies confirmed that the antibodies were specific for B. burgdorferi. Experimentally exposed dogs were asymptomatic. Naturally exposed dogs included both asymptomatic animals ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.4.648-653.1988
更新日期:1988-04-01 00:00:00
abstract::Several studies have identified thymidine excess in susceptibility test media as the cause of spurious resistance of various bacteria to sulfamethoxazole-trimethoprim. We document the phenomenon in Salmonella typhi and Salmonella paratyphi-A and demonstrate its occurrence in 3 of 17 (18%) lots of Mueller-Hinton agars ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.1.205-206.1986
更新日期:1986-01-01 00:00:00
abstract::We report the first case of spontaneous intrapartum Atopobium vaginae bacteremia identified by 16S rRNA gene sequencing. The bacterium was misidentified by RapID ANA II, API Rapid ID 32A, and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The likely source of bacteremia wa...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00212-12
更新日期:2012-07-01 00:00:00
abstract::Human adenovirus of strains subgenus F (AdV F) are the most common strains detected in acute gastroenteritis cases in developing countries. Subgenus F is represented by AdV serotype 40 (AdV-40) and AdV-41. Most of the reports have described the predominance of AdV-41 in acute gastroenteritis cases. To gain insight int...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01834-10
更新日期:2011-02-01 00:00:00
abstract::A serotyping scheme for Vibrio vulnificus predicated on the detection of lipopolysaccharide (LPS) antigens is proposed. The serovar O typing scheme used to type V. vulnificus employs polyclonal antisera raised in rabbits immunized with heat-killed whole-cell vaccines. Polyclonal typing sera produced in this manner cro...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.8.1684-1688.1991
更新日期:1991-08-01 00:00:00
abstract::The ability to isolate and propagate influenza virus is an essential tool for the yearly surveillance of circulating virus strains and to ensure accurate clinical diagnosis for appropriate treatment. The suitability of MDCK-SIAT1 cells, engineered to express increased levels of alpha-2,6-linked sialic acid receptors, ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00398-08
更新日期:2008-07-01 00:00:00
abstract::Since January 1996, and over a 3-year time span, a significant spread of serotype 6B multidrug-resistant (MDR) pneumococci, susceptible to penicillin and resistant to erythromycin, clindamycin, tetracycline, chloramphenicol, and trimethoprim-sulfamethoxazole, was noted in young carriers living in central and southern ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.2.581-585.2001
更新日期:2001-02-01 00:00:00
abstract::Rose Bengal and counterimmunoelectrophoresis, two tests that detect antibodies against different structural antigens, when carried out on spinal fluid permitted rapid diagnosis of human Brucella meningitis. The Rose Bengal test was positive in five out of five patients studied, and counterimmunoelectrophoresis was pos...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1978-02-01 00:00:00
abstract::The DNA fingerprint profiles and serotypes of 63 avian Pasteurella multocida field isolates, 13 attenuated vaccine isolates (propagated from vaccines manufactured by five companies), and 16 somatic reference strains were compared. DNA fingerprinting established the relationship of isolates that could not be distinguis...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.2.255-259.1993
更新日期:1993-02-01 00:00:00
abstract::Leifsonia aquatica is an aquatic bacterium that is typically found in environmental water habitats. Infections due to L. aquatica are rare and commonly catheter associated in immunocompromised patients. We report the first case of an acute septicemia caused by L. aquatica in a healthy immunocompetent host after cryope...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01339-13
更新日期:2013-11-01 00:00:00
abstract::Blastomyces dermatitidis and Histoplasma capsulatum are dimorphic fungi that often cause self-limited respiratory infections. However, they may also cause severe disseminated disease, depending on the level of the exposure to the organism and the host immune status. In addition, patients with infections caused by thes...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00673-11
更新日期:2011-09-01 00:00:00
abstract::Early identification of tuberculosis in the clinical setting is of great importance in order for specific therapy to be swiftly initiated. MB REDOX (Heipha Diagnostika), a growth-based medium without radioactive materials, was evaluated and was compared to the BACTEC system for detection of mycobacteria, including the...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.12.4048-4050.1999
更新日期:1999-12-01 00:00:00
abstract::This study shows that an unbiased amplification method applied to equine arteritis virus RNA significantly improves the sensitivity of the real-time reverse transcription-quantitative PCR (RT-qPCR) recommended by the World Organization for Animal Health. Twelve viral RNAs amplified using this method were hybridized on...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01935-14
更新日期:2015-01-01 00:00:00
abstract::Cryptococcus gattii emerged in North America in 1999 as a human and veterinary pathogen on Vancouver Island, British Columbia. The emergent subtype, VGIIa, and the closely related subtype VGIIb can now be found in the United States in Washington, Oregon, and California. We performed multilocus sequence typing and anti...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01505-09
更新日期:2010-02-01 00:00:00
abstract::Three forms of DNA-based typing procedures for Aspergillus fumigatus isolates have been developed over the last five years. The procedures are random amplification of polymorphic DNA (RAPD), restriction fragment length polymorphism (RFLP) detection, and Southern hybridizations with various repetitive sequence-based pr...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.1.87-93.1996
更新日期:1996-01-01 00:00:00
abstract::A quantitative, indirect, fluorescence immunoassay (FIAX; Whittaker Bioproducts, Inc.) was compared with the conventional indirect fluorescent-antibody test for detection of serum antibody to Borrelia burgdorferi. FIAX correlated well with the indirect fluorescent-antibody test (r = 0.72). FIAX is a convenient and dep...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.11.2218-2220.1987
更新日期:1987-11-01 00:00:00
abstract::The purpose of the investigation was to evaluate two commercially available identification systems: a new modification of the Staph-Zym system (Rosco, Tåstrup, Denmark) and the Staph ID 32 API system (API System, BioMérieux, Paris, France). A local standard method to be used in routine laboratories was also evaluated....
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.5.1150-1153.1995
更新日期:1995-05-01 00:00:00
abstract::The R/B and Minitek systems for the identification of Enterobacteriaceae were evaluated, and the results were compared with those obtained by conventional methods. Both systems were rapid and allowed correct identification of about 85% of the 294 isolates (23 species) examined. The individual biochemical reactions sho...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.1.6.504-508.1975
更新日期:1975-06-01 00:00:00
abstract::A colony immunoblot method was developed for serotyping the capsular polysaccharides expressed by Staphylococcus aureus isolates. The method was rapid and specific and was performed with either polyclonal or monoclonal antibodies specific for each of the capsule types. S. aureus isolates were obtained from patients wi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.28.12.2612-2615.1990
更新日期:1990-12-01 00:00:00