Abstract:
:Invasive pulmonary aspergillosis (IPA) is a frequently fatal infection in immunocompromised patients that is difficult to diagnose. Present methods for detection of Aspergillus spp. in bronchoalveolar lavage (BAL) fluid and in tissue vary in sensitivity and specificity. We therefore developed an A. fumigatus-specific quantitative real-time PCR-based assay utilizing fluorescent resonance energy transfer (FRET) technology. We compared the assay to quantitative culture of BAL fluid and lung tissue in a rabbit model of experimental IPA. Using an enzymatic and high-speed mechanical cell wall disruption protocol, DNA was extracted from samples of BAL fluid and lung tissues from noninfected and A. fumigatus-infected rabbits. A unique primer set amplified internal transcribed spacer regions (ITS) 1 and 2 of the rRNA operon. Amplicon was detected using FRET probes targeting a unique region of ITS1. Quantitation of A. fumigatus DNA was achieved by use of external standards. The presence of PCR inhibitors was determined by use of a unique control plasmid. The analytical sensitivity of the assay was =10 copies of target DNA. No cross-reactivity occurred with other medically important filamentous fungi. The assay results correlated with pulmonary fungal burden as determined by quantitative culture (r = 0.72, Spearman rank correlation; P = 0.0001). The mean number of genome equivalents detected in untreated animals was 3.86 log(10) (range, 0.86 to 6.39 log(10)) in tissue. There was a 3.53-log(10) mean reduction of A. fumigatus genome equivalents in animals treated with amphotericin B (AMB) (95% confidence interval, 3.38 to 3.69 log(10); P = 0.0001), which correlated with the reduction of residual fungal burden in lung tissue measured in terms of log(10) CFU/gram. The enhanced quantitative sensitivity of the real-time PCR assay was evidenced by detection of A. fumigatus genome in infarcted culture-negative lobes, by a greater number of mean genome equivalents compared to the number of CFU per gram in tissue and BAL fluid, and by superior detection of therapeutic response to AMB in BAL fluid compared to culture. This real-time PCR assay using FRET technology is highly sensitive and specific in detecting A. fumigatus DNA from BAL fluid and lung tissue in experimental IPA.
journal_name
J Clin Microbioljournal_title
Journal of clinical microbiologyauthors
O'Sullivan CE,Kasai M,Francesconi A,Petraitis V,Petraitiene R,Kelaher AM,Sarafandi AA,Walsh TJdoi
10.1128/jcm.41.12.5676-5682.2003subject
Has Abstractpub_date
2003-12-01 00:00:00pages
5676-82issue
12eissn
0095-1137issn
1098-660Xjournal_volume
41pub_type
杂志文章abstract::Sera from Brucella-infected bovines were analyzed by immunoblotting by using sonicated cell extracts of B. melitensis or B. abortus and a competitive enzyme-linked immunosorbent assay (ELISA) with monoclonal antibodies against outer membrane proteins (OMPs) with molecular masses of 10, 16.5, 19, 25 to 27, 36 to 38, an...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.12.3168-3174.1992
更新日期:1992-12-01 00:00:00
abstract::We compared the coreceptor tropism-predicting performance of a specific genotypic algorithm for HIV-1 subtype D and that of the geno2pheno algorithm with different cutoffs. The D-specific algorithm and geno2pheno with a false-positivity rate cutoff of 2.5% had the same concordance with the phenotypic determination. Th...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00798-13
更新日期:2013-09-01 00:00:00
abstract::We compared a nested PCR assay and microscopic examination of Giemsa-stained blood films for detection and identification of Plasmodium spp. in blood specimens. PCR was more sensitive than microscopy and capable of identifying malaria parasites at the species level when microscopy was equivocal. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.44.3.1087-1089.2006
更新日期:2006-03-01 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::Nocardia spp. can lead to local or disseminated infections, especially in immunocompromised patients. Combination therapy of amikacin and imipenem is commonly used to treat severe nocardial infections. We describe a patient with imipenem-resistant Nocardia cyriacigeorgica, which, to our knowledge, has not been previou...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02073-10
更新日期:2011-03-01 00:00:00
abstract::A 44-year-old diabetic female presented to a hospital in Jamaica with thermal burns. Trichosporon asahii was isolated from facial wounds, sputum, and a meningeal swab. Dissemination of the fungus was demonstrated in stained histological sections of the meninges and a brain abscess at autopsy. Pure growth of the fungus...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.05028-11
更新日期:2011-12-01 00:00:00
abstract::A trunk of human cytidylate-phosphate-deoxyguanylate-binding protein/CXXC finger protein 1 (CFP1), immobilized onto an aminohexyl-Sepharose column, can be used as a preanalytical tool for the selective enrichment of bacterial DNA from mixed solutions with high amounts of human background DNA for nucleic acid amplifica...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02242-08
更新日期:2009-04-01 00:00:00
abstract::A toxigenic Corynebacterium diphtheriae biovar mitis sequence type 136 (ST136) strain was recovered from a toe infection of an unvaccinated patient recently returned from India. The isolate was resistant to clindamycin, erythromycin (ermX positive), tetracycline, and trimethoprim-sulfamethoxazole, intermediate to ceft...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.05296-11
更新日期:2011-11-01 00:00:00
abstract::Transmissible spongiform encephalopathy strains can be differentiated by their behavior in bioassays and by molecular analyses of the disease-associated prion protein (PrP) in a posttranslationally transformed conformation (PrPSc). Until recently, isolates from cases of bovine spongiform encephalopathy (BSE) appeared ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00160-07
更新日期:2007-06-01 00:00:00
abstract::Forty-four nonreplicate clinical isolates of Pseudomonas aeruginosa that were resistant to extended-spectrum cephalosporins (ceftazidime and cefepime) and aztreonam, that putatively produced an acquired extended- spectrum beta-lactamase (ESBL), according to the results of a double-disk synergy test, and that had been ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.6.2523-2529.2004
更新日期:2004-06-01 00:00:00
abstract::Patients with chronic granulomatous disease have a marked defect in neutrophil oxidative metabolism and microbicidal activity. Asymptomatic mothers of males with the disease can usually be identified as heterozygous carriers by intermediate leukocyte function. Most mothers of females with the disease, however, have no...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.12.1.52-56.1980
更新日期:1980-07-01 00:00:00
abstract::An international program of surveillance of bloodstream infections (BSIs) in the United States, Canada, and South America between January and December 1997 detected 306 episodes of candidemia in 34 medical centers (22 in the United States, 6 in Canada, and 6 in South America). Eighty percent of the BSIs were nosocomia...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.36.7.1886-1889.1998
更新日期:1998-07-01 00:00:00
abstract::The test approved by the U.S. Food and Drug Administration for assessment of the barrier quality of medical exam gloves includes visual inspection and a water leak test. Neither method tests directly the ability of gloves to prevent penetration by microorganisms. Methods that use microorganisms (viruses and bacteria) ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.8.2725-2728.2002
更新日期:2002-08-01 00:00:00
abstract::A previously described sequence-based epidemiological typing method for clinical and environmental isolates of Legionella pneumophila serogroup 1 was extended by the investigation of three additional gene targets and modification of one of the previous targets. Excellent typeability, reproducibility, and epidemiologic...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.43.5.2047-2052.2005
更新日期:2005-05-01 00:00:00
abstract::Herpesviruses isolated from either domesticated or wild carnivores should be immunologically compared with known viruses of this group which could have been included in the diet of the animal before being considered to be previously undescribed herpesviruses native to the carnivore in question. ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.1.1.112-113.1975
更新日期:1975-01-01 00:00:00
abstract::Herpes simplex virus (HSV) causes acute and relapsing symptoms characterized by ulcerative lesions. Laboratory diagnosis of HSV in cutaneous or mucocutaneous lesions has historically been performed with the use of viral cell culture systems; however, these tests are laborious and suffer decreased sensitivity for advan...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00483-16
更新日期:2016-08-01 00:00:00
abstract::The long-chain fatty acids extracted from the whole cells of 12 clinically significant species of peptococci and peptostreptococci were characterized by gas-liquid chromatography. The resulting methylated fatty acid profiles (and some unidentified compounds) of 82 strains allowed the 12 species to be separated into fo...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:
更新日期:1976-12-01 00:00:00
abstract::Diagnostic laboratories are under increasing pressure to improve and expand their services. Greater flexibility in sample processing is a critical factor that can improve the time to results while reducing reagent waste, making laboratories more efficient and cost-effective. The introduction of the Abbott mPLUS featur...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.01672-13
更新日期:2013-12-01 00:00:00
abstract::In Brazil, the most common etiological agent of American tegumentary leishmaniasis is Leishmania (Viannia) braziliensis. In general, diagnostic techniques envisage the visualization of the parasite, but that technique has a low sensitivity. The main purpose of the present work was to evaluate the PCR as a routine tool...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.37.6.1819-1823.1999
更新日期:1999-06-01 00:00:00
abstract::An outbreak of folliculitis caused by Pseudomonas aeruginosa serotype O:7 occurred among the guests of a hotel in St. John's, Newfoundland, Canada, and the source of the infection was traced to the hotel whirlpool. Of 36 persons who used the whirlpool, 26 (72%) developed folliculitis within 1 to 5 days after exposure;...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章,评审
doi:10.1128/JCM.23.3.655-659.1986
更新日期:1986-03-01 00:00:00
abstract::Mucor ramosissimus Samutsevitsch is presented for the first time as an etiologic agent of cutaneous zygomycosis in a patient with aplastic anemia on immunosuppressive therapy. This report also represents the third case caused by this species reported in the literature. A biopsy taken from a lesion on the patient's thi...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.31.9.2523-2525.1993
更新日期:1993-09-01 00:00:00
abstract::A commercial DNA probe kit designed to detect rRNA from legionellae was evaluated for its ability to correctly discriminate between legionellae and non-legionellae taken from culture plates. The probe kit, made by the Gen-Probe Corp. (San Diego, Calif.), was radiolabeled with 125I, and probe bacterial RNA hybridizatio...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.23.3.481-484.1986
更新日期:1986-03-01 00:00:00
abstract::Clostridium hathewayi is a newly described Clostridium species isolated from the feces of healthy human individuals, but its clinical significance is not known. We describe a case of human infection associated with C. hathewayi. The bacterium (strain HKU18) was isolated from the blood culture of a 39-year-old patient ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.42.12.5947-5949.2004
更新日期:2004-12-01 00:00:00
abstract::Unbiased nontargeted metagenomic RNA sequencing (UMERS) has the advantage to detect known as well as unknown pathogens and, thus, can significantly improve the detection of viral, bacterial, parasitic, and fungal sequences in public health settings. In particular, conventional diagnostic methods successfully identify ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.02495-14
更新日期:2015-07-01 00:00:00
abstract::By using sequence analysis of Shiga toxin 1 (Stx 1) genes from human and ovine Stx-producing Escherichia coli (STEC) strains, we identified an Stx1 variant in STEC of human origin that was identical to the Stx1 variant from ovine STEC, but demonstrated only 97.1 and 96.6% amino acid sequence identity in its A and B su...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.40.4.1441-1446.2002
更新日期:2002-04-01 00:00:00
abstract::In this study we describe a multiplex PCR assay for the detection of nine clinically relevant antibiotic resistance genes of Staphylococcus aureus. Conditions were optimized to amplify fragments of mecA (encoding methicillin resistance), aacA-aphD (aminoglycoside resistance), tetK, tetM (tetracycline resistance), erm(...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.9.4089-4094.2003
更新日期:2003-09-01 00:00:00
abstract::Two multiplex PCRs were developed for the detection of necrotoxigenic Escherichia coli virulence genes. M1 contained the primers for the toxins and the aerobactin, and M2 contained the primers for the adhesins. They were validated by single PCRs performed with reference E. coli strains and by multiplex PCRs with necro...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/jcm.41.9.4480-4482.2003
更新日期:2003-09-01 00:00:00
abstract::The CO2, O2, and pH levels of commercially available blood culture bottles with tryptic soy broth medium from five different manufacturers were compared. Ranges of 1.3 to 6.9% for CO2, 1.1 to 6.0% for O2, and pH 6.94 to 7.26 were found. Different venting procedures revealed that blood culture bottles from which the ru...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.20.5.881-883.1984
更新日期:1984-11-01 00:00:00
abstract::Identification of members of the Mycobacterium tuberculosis complex and the M. avium-M. intracellulare complex (MAC) directly from primary BACTEC cultures was evaluated by using acridinium-ester-labeled DNA probes (AccuProbe; GenProbe, Inc., San Diego, Calif.). In preliminary experiments, blood present in samples was ...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.30.9.2427-2431.1992
更新日期:1992-09-01 00:00:00
abstract::Laboratory tests are essential for confirming sporadic cases and outbreaks of rubella. Detection of rubella virus is often necessary to confirm rubella cases and to identify specimens to be used to characterize wild-type rubella viruses. The sensitivities of four methods for detecting rubella virus infection using thr...
journal_title:Journal of clinical microbiology
pub_type: 杂志文章
doi:10.1128/JCM.00289-07
更新日期:2007-09-01 00:00:00