Abstract:
:The single-nucleotide variation 823C to T (His275Tyr), responsible for oseltamivir drug resistance has been detected in some isolates of the influenza A/H1N1/2009 virus. Early detection of the presence of this oseltamivir-resistant strain allows prompt consideration of alternative treatment options. An isolated-probe-asymmetric amplification PCR (Roche LightCycler v2.0) and high-resolution melting (HRM) method using unlabeled probes and amplified products (Idaho LightScanner 32) was designed and optimized to detect and estimate the proportion of H275Y mutants in influenza A/H1N1/2009 virus samples. The lower limit of quantification within the linear range of PCR assay detection was 200 copies/reaction. The melting peaks of the H275Y-specific unlabeled probe for the wild-type A/H1N1/2009 and H275Y mutant viruses were clearly distinguishable at 65.5°C and 69.0°C, respectively, at various ratios of wild-type/mutant virus population standards. The 95% detection limit for the 10% mutant sample pool was 1,200 copies/reaction (95% confidence interval, 669.7 to 3,032.6 copies/reaction). This HRM assay was tested with 116 archived clinical specimens. The quantitative HRM results obtained with samples containing mixed mutant-wild-type virus populations, at threshold cycle (C(T)) values of <29, compared well to those obtained with a pyrosequencing method performed by an independent laboratory. The quantitative feature of this assay allows the proportions of mutant and wild-type viral populations to be determined, which may assist in the conventional clinical management of infected patients and potentially more preemptive clinical management. This validated quantitative HRM method, with its low running cost, is well positioned as a rapid, high-throughput screening tool for oseltamivir resistance mutations in influenza A/H1N1/2009 virus-infected patients, with the potential to be adapted to other influenza virus species.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
Lee HK,Lee CK,Loh TP,Tang JW,Tambyah PA,Koay ESdoi
10.1128/JCM.01087-11subject
Has Abstractpub_date
2011-10-01 00:00:00pages
3555-9issue
10eissn
0095-1137issn
1098-660Xpii
JCM.01087-11journal_volume
49pub_type
杂志文章abstract::To determine sensitivity and specificity of immunofluorescence assay (IFA) for detection of Helicobacter pylori, we studied 151 patients. Biopsies of gastric mucosae were obtained for culture, histological testing, and IFA. H. pylori serum antibodies were tested by enzyme-linked immunosorbent assay. IFA was done on Fo...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.29.8.1748-1751.1991
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abstract::A case of severe progressive subcutaneous abscesses and necrotizing tenosynovitis of the right arm of a 30-year-old woman caused by Rhodococcus aurantiacus is reported. ...
journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.26.2.201-205.1988
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abstract::Mycoplasma hyopneumoniae is an important cause of pneumonia in pigs around the world, but confirming its presence in (or absence from) pigs can be difficult. Culture for diagnosis is impractical, and seroconversion is often delayed after natural infection, limiting the use of serology. Numerous PCR assays for the dete...
journal_title:Journal of clinical microbiology
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doi:10.1128/JCM.02366-07
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.15.6.1009-1015.1982
更新日期:1982-06-01 00:00:00
abstract::We examined clinical (pulmonary cryptococcosis and cryptococcal meningitis) and environmental (pigeon excreta) isolates of Cryptococcus neoformans var. neoformans (serotype A) in the southern Japanese prefecture of Nagasaki. The random amplified polymorphic DNA profiles obtained by using three primers revealed six pat...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.33.12.3328-3332.1995
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journal_title:Journal of clinical microbiology
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abstract::Two rapid diagnostic tests for Chlamydia trachomatis (Microtrak, Syva Co., Palo Alto, Calif.; and Chlamydiazyme, Abbott Laboratories, North Chicago, Ill.) were evaluated in comparison with growth of the organism in tissue culture for 2,030 urogenital specimens from men and women. Neither test performed as well as cult...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.25.10.1938-1943.1987
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01497-09
更新日期:2009-11-01 00:00:00
abstract::An enzyme-linked immunosorbent assay for the detection of antibodies to dengue virus is described. This method correlates well with a hemagglutination inhibition technique. The enzyme-linked immunosorbent assay can also be specific for human immunoglobulin M antibodies when a mu-chain-specific antiglobulin-enzyme conj...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
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abstract::Influenza epidemics arise through the accumulation of viral genetic changes, culminating in a novel antigenic type that is able to escape host immunity. Following an outbreak of the A/Fujian/411/2002-like strains in Asia, including China, Japan, and South Korea, in 2002, Australia and New Zealand experienced substanti...
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.6.1257-1261.1989
更新日期:1989-06-01 00:00:00
abstract::INNO-LiPA and PapType human papillomavirus (HPV) genotyping assays were compared for detection of HPV genotypes on archival vulvar tissue. The INNO-LiPA assay detected 49 HPV-16 infections, compared with 47 detected by the PapType assay. The INNO-LiPA assay detected amplifiable DNA in 59 (91%) biopsy specimens, compar...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00516-11
更新日期:2011-11-01 00:00:00
abstract::We isolated Shiga toxin-producing Escherichia coli O157:H7 strains resistant to third-generation cephalosporins. The resistant strains harbored blaCMY-2, a plasmid-mediated AmpC β-lactamase. Genotyping of isolates revealed the possible spread of this problematic bacterium. Results suggested the importance of the inves...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01263-15
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01958-18
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00498-14
更新日期:2014-07-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.2.179-184.1993
更新日期:1993-02-01 00:00:00
abstract::By use of the nearly perfectly colinear genomes of Rickettsia conorii and Rickettsia prowazekii, we compared the usefulness of three types of sequences for typing of R. conorii isolates: (i) 5 variable coding genes comprising the 16S ribosomal DNA, gltA, ompB, and sca4 (gene D) genes, which are present in both genomes...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.12.5757-5766.2004
更新日期:2004-12-01 00:00:00
abstract::Inactivation of penicillin and gentamicin in cultures was achieved by using monoclonal antibodies against these antibiotics. A viridans group streptococcus (penicillin MIC, less than or equal to 0.06 micrograms/ml) and Escherichia coli ATCC 35218 (gentamicin MIC, less than or equal to 1 microgram/ml) were able to grow...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.26.9.1904-1906.1988
更新日期:1988-09-01 00:00:00
abstract::Mycobacterium avium subsp. paratuberculosis causes Johne's disease, a systemic infection and chronic inflammation of the intestine that affects many species, including primates. Infection is widespread in livestock, and human populations are exposed. Johne's disease is associated with immune dysregulation, with involv...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01371-07
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.27.3.580-582.1989
更新日期:1989-03-01 00:00:00
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journal_title:Journal of clinical microbiology
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.12.4423-4427.2002
更新日期:2002-12-01 00:00:00
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journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.5.2068-2079.2003
更新日期:2003-05-01 00:00:00
abstract::Jena virus (JV) is a bovine enteric calicivirus that causes diarrhea in calves. The virus is approximately 30 nm in diameter and has a surface morphology similar to the human Norwalk virus. The genome sequence of JV was recently described, and the virus has been assigned to the genus Norovirus of the family CALICIVIRI...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.6.2300-2305.2003
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abstract::Bordetella bronchiseptica and toxigenic Pasteurella multocida are the etiologic agents of swine atrophic rhinitis. Methods currently used for their identification are time-consuming and suffer from a lack of sensitivity. We describe a colony lift-hybridization assay for detection of B. bronchiseptica and toxigenic P. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.11.3342-3346.1998
更新日期:1998-11-01 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::Spontaneously occurring urinary tract infection (UTI) in dogs was exploited as an experiment of nature to gain insights into UTI pathogenesis in humans. Concurrent urinary and rectal Escherichia coli isolates from 37 dogs with UTI were compared with respect to phylogenetic background, O antigens, and extended virulenc...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.1.337-345.2003
更新日期:2003-01-01 00:00:00
abstract::Ventilator-associated pneumonia is frequently diagnosed with quantitative cultures of samples obtained by bronchoscopic techniques, a method associated with high costs and potential adverse effects. Quantitative cultures of endotracheal aspirates are easier and cheaper to obtain, and good correlations between the resu...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.35.3.796-798.1997
更新日期:1997-03-01 00:00:00
abstract::The development and evaluation of a new selective medium (colistin-oxolinic acid-blood agar) for streptococci is described. Streptococci of medical and veterinary importance grew well on the medium. Gram-negative organisms, staphylococci. Bacillus spp., and coryneforms are all inhibited. It was concluded that the medi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.19.1.4-7.1984
更新日期:1984-01-01 00:00:00