Abstract:
:A quantitative PCR assay for the detection of human herpesvirus 6 (HHV-6) (variants A and B) and HHV-7 DNAs in clinical samples was developed. The assay uses a nonhomologous internal standard (IS) for each virus that is coamplified with the wild-type target sequence in the same vial and with the same pair of primers. This method allows for a correction of the variability of efficiency of the PCR technique. A standard curve is constructed for each experiment by coamplification of known quantities of the cloned HHV-6 or HHV-7 target templates with the respective IS. Absolute quantitation of the test samples is then achieved by determining the viral target/IS ratio of the hybridization signals of the amplification products and plotting this value against the standard curve. Using this assay, we quantitated the amount of HHV-6 or HHV-7 DNA in infected cell cultures and demonstrated an inhibitory effect of phosphonoformic acid on the replication of HHV-6 and HHV-7 in vitro. As the first clinical application of this procedure, we performed preliminary measurements of the loads of HHV-6 and HHV-7 in lymph nodes from patients with Hodgkin's disease and AIDS. Application of this quantitative PCR method should be helpful for elucidating the pathogenic roles of HHV-6 and HHV-7.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Secchiero P,Zella D,Crowley RW,Gallo RC,Lusso Pdoi
10.1128/JCM.33.8.2124-2130.1995subject
Has Abstractpub_date
1995-08-01 00:00:00pages
2124-30issue
8eissn
0095-1137issn
1098-660Xjournal_volume
33pub_type
杂志文章abstract::The nucleic acid sequence of the putative 5'-untranslated (5PUT) region of hepatitis C virus (HCV), determined for samples obtained from a variety of geographic origins, was found to be over 98% conserved among all isolates. On the basis of this signature sequence for HCV, a viral RNA assay was developed by using cDNA...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.11.2528-2534.1991
更新日期:1991-11-01 00:00:00
abstract::The zygomycetous mold Cokeromyces recurvatus was isolated from the pleural and peritoneal fluids of a 64-year-old man with a history of peptic ulcer disease and alcohol abuse. The patient presented to the hospital with severe abdominal pain and a ruptured duodenal ulcer. This is one of the few times that C.recurvatus ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.10.2601-2603.1996
更新日期:1996-10-01 00:00:00
abstract::Herpes simplex virus (HSV) infections are very common in the general population and among immunocompromised patients. Acyclovir (ACV) is an effective treatment which is widely used. We deemed it essential to conduct a wide and coordinated survey of the emergence of ACV-resistant HSV strains. We have formed a network o...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.1.242-249.2004
更新日期:2004-01-01 00:00:00
abstract::We retrospectively compared the utility of a fungal isolation device (Isolator) versus conventional techniques for recovering fungal organisms from blood cultures obtained from neutropenic cancer patients. Positive cultures were deemed true pathogens, possible pathogens, or contaminants according to laboratory and cli...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.1.290-293.1998
更新日期:1998-01-01 00:00:00
abstract::We describe two cases in Brazil of human subcutaneous infections due to Phaeoacremonium spp. The first case was caused by Phaeoacremonium aleophilum. The patient presented with a unique fistulized nodule on the left ankle. The fungus was detected by direct microscopic examination and was isolated repeatedly from mater...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.3.1332-1336.2003
更新日期:2003-03-01 00:00:00
abstract::HIV-testing algorithms for preexposure prophylaxis (PrEP) should be optimized to minimize the risk of drug resistance, the time off PrEP required to evaluate false-positive screening results, and costs and to expedite the start of therapy for those confirmed to be infected. HIV rapid tests (RTs) for anti-HIV antibodie...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,多中心研究
doi:10.1128/JCM.01540-14
更新日期:2015-01-01 00:00:00
abstract::In recent years testing responsibilities for high-consequence pathogens have been expanded from national reference laboratories into networks of local and regional laboratories in order to support enhanced disease surveillance and to test for surge capacity. This movement of testing of select agents and high-consequen...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.44.1.67-70.2006
更新日期:2006-01-01 00:00:00
abstract::Herpes simplex virus (HSV) infections of the central nervous system (CNS) are associated with significant morbidity and mortality rates in children. This study assessed the impact of a direct HSV (dHSV) PCR assay on the time to result reporting and the duration of acyclovir therapy for children with signs and symptoms...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02559-16
更新日期:2017-05-01 00:00:00
abstract::Screening for Chlamydia trachomatis infection in men has traditionally been limited to men who present with urethral symptoms, thereby limiting the detection of asymptomatic chlamydia infection in men. In order to effectively screen both symptomatic and asymptomatic men, we evaluated a newly developed polymerase chain...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.5.1209-1212.1993
更新日期:1993-05-01 00:00:00
abstract::Clonal spread and horizontal transfer in the spread of vancomycin resistance genes were investigated. Multiplex PCR, pulsed-field gel electrophoresis (PFGE), hybridization of enterococcal plasmids with the vanA and vanB probes, and sequencing of a fragment of vanB were used in the analysis. Before May 1996, 12 vancomy...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.12.3934-3939.1999
更新日期:1999-12-01 00:00:00
abstract::Several problems remain before molecular biology-based techniques, such as PCR, are widely accepted for the detection of infectious agents. Among the most formidable of these problems are the inability of the tests to distinguish between viable and nonviable organisms. We approached this problem by using the fact that...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.5.1161-1165.1997
更新日期:1997-05-01 00:00:00
abstract::An automated RNA isolation procedure on the Qiagen BioRobot is described for performing viral load tests for human immunodeficiency virus (HIV) with the Amplicor HIV type 1 test. The new procedure improves the precision of the assay and requires significantly less labor than the presently used manual RNA isolation pro...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.1.439-440.2004
更新日期:2004-01-01 00:00:00
abstract::In a prospective study, 139 serial blood samples from 15 transplant recipients were assessed for the presence of cytomegalovirus (CMV) by virus isolation (CMV viremia) and by direct staining of CMV antigens (CMV Ag) in blood leukocytes (CMV antigenemia). CMV was isolated from 23 samples, whereas CMV Ag was detected in...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.12.2531-2535.1988
更新日期:1988-12-01 00:00:00
abstract::An in situ DNA hybridization kit for cytomegalovirus (CMV) was evaluated for the detection of CMV in centrifugation culture. Of 61 clinical specimens, 17 (27.8%) were positive for CMV by monoclonal antibody staining following centrifugation. Of the 17 positive specimens, 15 were detected by DNA hybridization (24.5%). ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.1.21-23.1989
更新日期:1989-01-01 00:00:00
abstract::Two high-copy-number insertion sequences, IS2404 and IS2606, were recently identified in Mycobacterium ulcerans and were shown by Southern hybridization to possess restriction fragment length polymorphism between strains from different geographic origins. We have designed a simple genotyping method that captures these...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.38.4.1482-1487.2000
更新日期:2000-04-01 00:00:00
abstract::While high-copy-number IS6110-based restriction fragment length polymorphism (HCN-RFLP) is the gold standard for typing most Mycobacterium tuberculosis strains, the time taken for culturing and low throughput make it impractical for large-scale prospective typing of large numbers of isolates. The development of a new ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01428-05
更新日期:2006-07-01 00:00:00
abstract::Using the Staphylococcus aureus absorption method, we have modified a commercial radioimmunoassay kit for the determination of total antibody to hepatitis B core antigen so that we can now determine the predominant globulin species of that antibody. One-hundred percent of acute hepatitis B sera had a predominance of i...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.16.5.973-975.1982
更新日期:1982-11-01 00:00:00
abstract::The etiology of Crohn's disease in humans is largely unknown. Clinical signs of Crohn's disease partly resemble the clinical picture of Johne's disease in ruminants caused by Mycobacterium avium subsp. paratuberculosis. Because of the high prevalence of these bacteria in (products of) ruminants and their remarkable th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.11.5022-5028.2004
更新日期:2004-11-01 00:00:00
abstract::Bilophila wadsworthia is a common inhabitant of the human colon and has been associated with appendicitis and other local sites of inflammation in humans. Challenge-exposure or prevalence studies in laboratory and other animals have not been reported. B. wadsworthia is closely related phylogenetically to Desulfovibrio...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.39.4.1577-1579.2001
更新日期:2001-04-01 00:00:00
abstract::Clinical evidence for failure with beta-lactam therapy has been lacking for patients with borderline oxacillin-resistant Staphylococcus aureus (BORSA) infections. We describe a failure of cloxacillin for a patient with endocarditis due to BORSA. The isolate also had false-negative thermonuclease and coagulase test res...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00571-08
更新日期:2009-03-01 00:00:00
abstract::Nosocomial vancomycin-resistant Enterococcus (VRE) infections have been described in only small numbers of pediatric patients. In none of these studies were multivariate analyses performed to assess which factors were independent risk factors in these patients. In the present cohort study of patients admitted to our h...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.2.413-416.1999
更新日期:1999-02-01 00:00:00
abstract::A set of universal degenerate primers which amplified, by PCR, a 600-bp oligomer encoding a portion of the 60-kDa heat shock protein (HSP60) of both Staphylococcus aureus and Staphylococcus epidermidis were developed. However, when used as a DNA probe, the 600-bp PCR product generated from S. epidermidis failed to cro...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.34.4.818-823.1996
更新日期:1996-04-01 00:00:00
abstract::Pulsed-field gel electrophoresis (PFGE) of XbaI-digested chromosomal DNA was performed on 133 strains of Salmonella enterica serovar Typhi obtained from Papua New Guinea, with the objective of assessing the temporal variation of these strains. Fifty-two strains that were isolated in 1992 and 1994 were of one phage typ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.11.4156-4160.2002
更新日期:2002-11-01 00:00:00
abstract::In this study, we determined the sequence of the EcoRI-PstI fragment of the plasmid pCVD432, also termed the enteroaggregative Escherichia coli (EAggEC) probe. A primer pair complementary to this probe was designed for PCR amplification of a 630-bp region. Comparison of the analysis of the EAggEC probe sequence with t...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.3.701-705.1995
更新日期:1995-03-01 00:00:00
abstract::Many central vascular catheters (CVCs) are removed unnecessarily because current diagnostic methods for CVC-associated infection are unreliable. A quantitative PCR assay using primers and probe targeted to bacterial 16S ribosomal DNA was used to measure the levels of bacterial DNA in blood samples drawn through the CV...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.42.4.1402-1408.2004
更新日期:2004-04-01 00:00:00
abstract::Buruli ulcer, an infection caused by Mycobacterium ulcerans, is, after tuberculosis and leprosy, the third most common mycobacterial disease. The mode of transmission of M. ulcerans is not exactly known, but since Buruli ulcer often occurs in focalized swampy areas, it is assumed that there is a reservoir of the patho...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.9.4231-4237.2003
更新日期:2003-09-01 00:00:00
abstract::A selective agar medium for isolation of virulent Yersinia enterocolitica (VYE agar) was developed for the rapid and accurate isolation of virulent Y. enterocolitica from environmental samples highly contaminated with environmental Yersinia organisms, as well as for isolation from clinical specimens. VYE agar provided...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.6.1068-1073.1987
更新日期:1987-06-01 00:00:00
abstract::Hybridomas secreting specific monoclonal antibodies (MAb) to all members of the genus Leptospira (clone LF9) and those that are specific only to the pathogenic species (clones LD5 and LE1) were produced. MAb LF9, which was immunoglobulin G1 (IgG1), reacted to a 38-kDa component of the sodium dodecyl sulfate-polyacryla...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.3.480-489.2002
更新日期:2002-02-01 00:00:00
abstract::We report an outbreak of Saccharomyces cerevisiae subtype boulardii fungemia among three intensive care unit roommates of patients receiving lyophilized preparations of this fungus. The fungemia was probably due to central venous catheter contamination and resolved after fluconazole treatment. The need for stringent a...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
doi:10.1128/jcm.41.11.5340-5343.2003
更新日期:2003-11-01 00:00:00
abstract::Coxsackievirus (CV) strains CVA2, CVA4, CVA5, and CVA10 were isolated from patients with hand, foot, and mouth disease during a 2009 outbreak in China. Full genome sequences for four representative strains, CVA2/SD/CHN/09 (A2SD09), CVA4/SZ/CHN/09 (A4SZ09), CVA5/SD/CHN/09 (A5SD09), and CVA10/SD/CHN/09 (A10SD09), were d...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00007-11
更新日期:2011-07-01 00:00:00