Application of 16S rRNA gene PCR to study bowel flora of preterm infants with and without necrotizing enterocolitis.

Abstract:

:The purpose of the present study was to determine the extent to which bacteria not detected by culture contribute to the microbial flora of the bowel of preterm infants with and without neonatal necrotizing enterocolitis (NEC). Fecal samples from 32 preterm infants in special care baby units including samples from 10 infants with NEC were examined by culture and PCR amplification of the 16S rRNA gene (rDNA). The 16S rDNA V3 region was amplified with eubacterial primers, and the amplification products derived from the fecal sample DNA were compared with the products from individual cultured isolates by PCR and denaturing gradient gel electrophoresis (PCR-DGGE), allowing the DNA from uncultured bacteria to be identified. For the 22 infants without NEC weekly samples were examined for a mean of 5.3 postnatal weeks. The total number of types detected by culture combined with PCR-DGGE was 10.1 per infant, of which PCR-DGGE contributed 10.4% of the types identified. Additional types detected by PCR-DGGE were found in 14 (63.6%) of the 22 infants. The majority of the sequences associated with uncultured bacteria showed > 90% 16S rDNA sequence identity with sequences from culturable human enteric flora, and all were found in single infants with the exception of sequences indistinguishable by DGGE from seven infants. These sequences showed > 90% sequence identity with the 16S rDNA of Streptococcus salivarius and may have been derived from upper gastrointestinal or respiratory tract flora. In the present study uncultured bacteria detected by PCR-DGGE were no more frequent in fecal samples from infants with NEC than in samples from infants without NEC, although these findings do not exclude the possibility of unrecognized bacteria associated with the mucosa of the small intestine of infants with NEC.

journal_name

J Clin Microbiol

authors

Millar MR,Linton CJ,Cade A,Glancy D,Hall M,Jalal H

doi

10.1128/JCM.34.10.2506-2510.1996

subject

Has Abstract

pub_date

1996-10-01 00:00:00

pages

2506-10

issue

10

eissn

0095-1137

issn

1098-660X

journal_volume

34

pub_type

杂志文章
  • Use of new T-cell-based cell lines expressing two luciferase reporters for accurately evaluating susceptibility to anti-human immunodeficiency virus type 1 drugs.

    abstract::Two new T-cell-based reporter cell lines were established to measure human immunodeficiency virus type 1 (HIV-1) infectivity. One cell line naturally expresses CD4 and CXCR4, making it susceptible to X4-tropic viruses, and the other cell line, in which a CCR5 expression vector was introduced, is susceptible to both X4...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01708-06

    authors: Chiba-Mizutani T,Miura H,Matsuda M,Matsuda Z,Yokomaku Y,Miyauchi K,Nishizawa M,Yamamoto N,Sugiura W

    更新日期:2007-02-01 00:00:00

  • Molecular epidemiology of Aspergillus fumigatus: an in-depth genotypic analysis of isolates involved in an outbreak of invasive aspergillosis.

    abstract::We recently reported an outbreak of invasive aspergillosis in the major heart surgery unit of Hospital Gregorio Marañón, Madrid, Spain (T. Peláez, P. Muñoz, J. Guinea, M. Valerio, M. Giannella, C. H. W. Klaassen, and E. Bouza, Clin. Infect. Dis., in press). Aspergillus fumigatus was isolated from clinical samples from...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01159-11

    authors: Guinea J,García de Viedma D,Peláez T,Escribano P,Muñoz P,Meis JF,Klaassen CH,Bouza E

    更新日期:2011-10-01 00:00:00

  • Molecular analysis of microflora associated with dentoalveolar abscesses.

    abstract::The microflora associated with three dentoalveolar abscesses was determined by cultural and molecular methods. 16S rRNA genes were randomly amplified by means of conserved eubacterial primers and cloned. Restriction fragment length polymorphism analysis of the clones and amplified genes encoding 16S rRNA from the cult...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.34.3.537-542.1996

    authors: Dymock D,Weightman AJ,Scully C,Wade WG

    更新日期:1996-03-01 00:00:00

  • A Multiplexed Serologic Test for Diagnosis of Lyme Disease for Point-of-Care Use.

    abstract::Single multiplexed assays could replace the standard 2-tiered (STT) algorithm recommended for the laboratory diagnosis of Lyme disease if they perform with a specificity and a sensitivity superior or equal to those of the STT algorithm. We used human serum rigorously characterized to be sera from patients with acute- ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.01142-19

    authors: Arumugam S,Nayak S,Williams T,di Santa Maria FS,Guedes MS,Chaves RC,Linder V,Marques AR,Horn EJ,Wong SJ,Sia SK,Gomes-Solecki M

    更新日期:2019-11-22 00:00:00

  • Drug-associated resistance mutations in plasma and peripheral blood mononuclear cells of human immunodeficiency virus type 1-infected patients for whom highly active antiretroviral therapy is failing.

    abstract::In 32 patients for whom highly active antiretroviral therapy was failing, a good agreement between drug resistance-associated mutations in plasma and peripheral blood mononuclear cells (PBMCs) was found (k = 0.85). The mutations with the lowest agreement were 20R, 63P, and 84V in the protease gene and 184V in the reve...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.4.1760-1762.2003

    authors: Sarmati L,Nicastri E,Uccella I,D'Ettorre G,Parisi SG,Palmisano L,Galluzzo C,Concia E,Vullo V,Vella S,Andreoni M

    更新日期:2003-04-01 00:00:00

  • Detection of Legionella pneumophila capsular-like envelope antigens by counterimmunoelectrophoresis.

    abstract::The capsular-like envelope of Legionella pneumophila strains Togus 1 (serotype 2) and Philadelphia 1 (serotype 1) was isolated and purified by column chromatography on Sepharose 6B. Antibody raised in rabbits to these two antigenic materials did not cross-react in gel diffusion. Upon electrophoresis followed by gel di...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.13.4.637-642.1981

    authors: Smith RA,DiGiorgio S,Darner J,Wilhelm A

    更新日期:1981-04-01 00:00:00

  • Detection of Mycoplasma pneumoniae by real-time nucleic acid sequence-based amplification.

    abstract::Real-time isothermal nucleic acid sequence-based amplification (RT-NASBA) was applied to the detection of Mycoplasma pneumoniae. In vitro-generated M. pneumoniae RNA was used to assess the sensitivity of the assay. The 95% hit rate was 148 molecules of M. pneumoniae RNA in the amplification and 10(4) molecules of in v...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.41.9.4448-4450.2003

    authors: Loens K,Ieven M,Ursi D,Beck T,Overdijk M,Sillekens P,Goossens H

    更新日期:2003-09-01 00:00:00

  • PCR ribotyping and arbitrarily primed PCR for typing strains of Clostridium difficile from a Polish maternity hospital.

    abstract::Detection of the source of Clostridium difficile strains is of importance for the control of the nosocomial spread of this microorganism. For this purpose, vaginal and rectal swabs from 183 mothers, duplicate fecal samples (taken on days 1 and 4 after birth) from 183 neonates, and 94 environmental samples were culture...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.8.2016-2021.1995

    authors: Martirosian G,Kuipers S,Verbrugh H,van Belkum A,Meisel-Mikolajczyk F

    更新日期:1995-08-01 00:00:00

  • Mycobacterium avium complex, an emerging pathogen in Massachusetts.

    abstract::We report a study of 1,953 patients whose laboratory records from 1972 through 1983 at the Massachusetts Mycobacteria Reference Laboratory indicated the isolation of Mycobacterium avium complex (MAC) organisms. At least one clinical specimen from each patient during this period exhibited the organism. The incidence of...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.22.1.9-12.1985

    authors: du Moulin GC,Sherman IH,Hoaglin DC,Stottmeier KD

    更新日期:1985-07-01 00:00:00

  • Aberrant forms of Escherichia coli in blood cultures: in vitro reproduction of an in vivo observation.

    abstract::Aberrant filamentous forms of Escherichia coli were observed on direct Gram stain of blood cultures from a patient being treated with the beta-lactam antibiotic cephalexin. After the institution of an alternative antibiotic regimen which included a different cell wall-active agent, E. coli of normal morphology was det...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.12.5.690-694.1980

    authors: Magnussen CR,Hruska JF

    更新日期:1980-11-01 00:00:00

  • Detection of PCR inhibitors in cervical specimens by using the AMPLICOR Chlamydia trachomatis assay.

    abstract::To determine that susceptibility of AMPLICOR Chlamydia trachomatis PCR to inhibitory factors possibly present in cervical specimens, we obtained cervical specimens from 200 gynecology patients attending our outpatient clinic. The prevalence of C. trachomatis infection was 4.1%, as determined by cell culture. All AMPLI...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.34.12.3072-3074.1996

    authors: Verkooyen RP,Luijendijk A,Huisman WM,Goessens WH,Kluytmans JA,van Rijsoort-Vos JH,Verbrugh HA

    更新日期:1996-12-01 00:00:00

  • Characterization of Clostridium difficile strains isolated from patients in Ontario, Canada, from 2004 to 2006.

    abstract::Clostridium difficile is the bacterium most commonly surmised to cause antimicrobial- and hospital-associated diarrhea in developed countries worldwide, and such infections are thought to be increasing in frequency and severity. A laboratory-based study was carried out to characterize C. difficile strains isolated fro...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02437-07

    authors: Martin H,Willey B,Low DE,Staempfli HR,McGeer A,Boerlin P,Mulvey M,Weese JS

    更新日期:2008-09-01 00:00:00

  • Investigation of apparent false-positive urine latex particle agglutination tests for the detection of group B streptococcus antigen.

    abstract::In our nursery, we identified neonates with positive urine latex particle agglutination (LPA) tests for group B streptococcus (GBS) antigen who did not have corroborating cultural evidence of infection. To investigate the mechanisms underlying these apparent false-positive reactions, we examined the urine LPA test in ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.27.10.2214-2217.1989

    authors: Harris MC,Deuber C,Polin RA,Nachamkin I

    更新日期:1989-10-01 00:00:00

  • Chromosomal DNA fingerprint patterns produced with IS6110 as strain-specific markers for epidemiologic study of tuberculosis.

    abstract::Mycobacterium tuberculosis isolates were studied by comparing chromosomal DNA fingerprint patterns produced by digestion of chromosomal DNA with BamHI, followed by agarose electrophoresis and hybridization with radiolabeled probes of insertion sequence IS6110. DNA fingerprints of 14 isolates from separate members of f...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.29.9.2030-2033.1991

    authors: Mazurek GH,Cave MD,Eisenach KD,Wallace RJ Jr,Bates JH,Crawford JT

    更新日期:1991-09-01 00:00:00

  • Malnutrition Is Associated with Protection from Rotavirus Diarrhea: Evidence from a Longitudinal Birth Cohort Study in Bangladesh.

    abstract::Rotavirus is a leading cause of dehydrating diarrhea and death among infants and children globally, particularly in communities of the developing world. While numerous studies have described the complex relationships among infectious diarrhea, growth faltering, and poverty, the impact of nutritional status on suscepti...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00916-16

    authors: Verkerke H,Sobuz S,Ma JZ,Petri SE,Reichman D,Qadri F,Rahman M,Haque R,Petri WA Jr

    更新日期:2016-10-01 00:00:00

  • Predicting susceptibility of Streptococcus pneumoniae to ceftriaxone and cefotaxime by cefuroxime and ceftizoxime disk diffusion testing.

    abstract::In this study, disk diffusion testing with ceftizoxime and cefuroxime was evaluated for use in predicting the susceptibility of Streptococcus pneumoniae to ceftriaxone and cefotaxime. Of the 194 isolates included in this study, 138 were susceptible, 34 were intermediate, and 22 were resistant to cefotaxime by MIC test...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.37.11.3707-3710.1999

    authors: Williams-Bouyer N,Hernandez A,Reisner BS

    更新日期:1999-11-01 00:00:00

  • Detection and multigenic characterization of a herpesvirus associated with malignant catarrhal fever in white-tailed deer (Odocoileus virginianus) from Missouri.

    abstract::Between 1998 and 2001, tissues from four captive white-tailed deer were observed to have histologic lesions of systemic lymphocytic vasculitis. These lesions suggested malignant catarrhal fever, although epizootic hemorrhagic disease and bluetongue were included in the differential diagnosis. Initial diagnostic effort...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.40.4.1311-1318.2002

    authors: Kleiboeker SB,Miller MA,Schommer SK,Ramos-Vara JA,Boucher M,Turnquist SE

    更新日期:2002-04-01 00:00:00

  • Isolation of enterotoxigenic Bacteroides fragilis from Bangladeshi children with diarrhea: a controlled study.

    abstract::We undertook a controlled study of children younger than 5 years in Bangladesh to determine whether enterotoxigenic Bacteroides fragilis (ETBF) was associated with diarrhea in this population. ETBF was isolated from 22 (6.1%) of 358 patients and 5 (1.2%) of 425 controls (P = 0.0001). In children younger than 1 year, h...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.32.4.960-963.1994

    authors: Sack RB,Albert MJ,Alam K,Neogi PK,Akbar MS

    更新日期:1994-04-01 00:00:00

  • Evaluation of Clostridium difficile fecal load and limit of detection during a prospective comparison of two molecular tests, the illumigene C. difficile and Xpert C. difficile/Epi tests.

    abstract::In a large prospective comparison, the illumigene test detected Clostridium difficile in 98% of toxin-positive and 58% of toxin-negative samples confirmed positive by other methods. The Xpert was uniformly sensitive. Most samples with discrepant results had C. difficile concentrations below the illumigene limit of det...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.02120-12

    authors: Gyorke CE,Wang S,Leslie JL,Cohen SH,Solnick JV,Polage CR

    更新日期:2013-01-01 00:00:00

  • PCR strategy for identification and differentiation of small pox and other orthopoxviruses.

    abstract::Rapid identification and differentiation of orthopoxviruses by PCR were achieved with primers based on genome sequences encoding the hemagglutinin (HA) protein, an infected-cell membrane antigen that distinguishes orthopoxviruses from other poxvirus genera. The initial identification step used a primer pair of consens...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.33.8.2069-2076.1995

    authors: Ropp SL,Jin Q,Knight JC,Massung RF,Esposito JJ

    更新日期:1995-08-01 00:00:00

  • Comparison between the Hybrid Capture II Test and an SPF1/GP6+ PCR-based assay for detection of human papillomavirus DNA in cervical swab samples.

    abstract::We compared the efficacy of human papillomavirus (HPV) DNA detection between a PCR-based genechip (Easychip HPV Blot [hereafter referred to as HPV Blot]; King Car, Taiwan) method and Hybrid Capture II (HCII; Digene, Gaithersburg, MD) in women with previous normal (n = 146) or abnormal (> or =atypical squamous cells of...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.44.5.1733-1739.2006

    authors: Huang SL,Chao A,Hsueh S,Chao FY,Huang CC,Yang JE,Lin CY,Yan CC,Chou HH,Huang KG,Huang HJ,Wu TI,Tseng MJ,Qiu JT,Lin CT,Chang TC,Lai CH

    更新日期:2006-05-01 00:00:00

  • Molecular follow-up of Salmonella enterica subsp. enterica serovar Agona infection in cattle and humans.

    abstract::Salmonella enterica subsp. enterica serovar Agona was not frequently encountered in Finland until an increase in rates of isolation among animal and feed was seen in 1994. A small outbreak among cattle farms in the regions of Oulu and Vaasa in northwestern Finland in 1994-1995 included eight farms. After the outbreak,...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/jcm.40.10.3648-3653.2002

    authors: Lindqvist N,Siitonen A,Pelkonen S

    更新日期:2002-10-01 00:00:00

  • Genetic heterogeneity of Borrelia burgdorferi sensu lato in the southern United States based on restriction fragment length polymorphism and sequence analysis.

    abstract::Fifty-six strains of Borrelia burgdorferi sensu lato, isolated from ticks and vertebrate animals in Missouri, South Carolina, Georgia, Florida, and Texas, were identified and characterized by PCR-restriction fragment length polymorphism (RFLP) analysis of rrf (5S)-rrl (23S) intergenic spacer amplicons. A total of 241 ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.39.7.2500-2507.2001

    authors: Lin T,Oliver JH Jr,Gao L,Kollars TM Jr,Clark KL

    更新日期:2001-07-01 00:00:00

  • Serratia rubidaea as an invasive pathogen.

    abstract::Serratia rubidaea biotype 1 was isolated from the bile and blood of a patient with a bile tract carcinoma obstructing the common bile duct and who underwent invasive procedures. The infection was cleared after adequate treatment with antibiotics. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.34.1.216-217.1996

    authors: Ursua PR,Unzaga MJ,Melero P,Iturburu I,Ezpeleta C,Cisterna R

    更新日期:1996-01-01 00:00:00

  • Novel approach to in vitro drug susceptibility assessment of clinical strains of Leishmania spp.

    abstract::Resistance to antimonial drugs has been documented in Leishmania isolates transmitted in South America, Europe, and Asia. The frequency and distribution of resistance to these and other antileishmanial drugs are unknown. Technical constraints have limited the assessment of drug susceptibility of clinical strains of Le...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.00216-12

    authors: Fernández O,Diaz-Toro Y,Valderrama L,Ovalle C,Valderrama M,Castillo H,Perez M,Saravia NG

    更新日期:2012-07-01 00:00:00

  • Evaluation of a radial immunodiffusion test with polysaccharide B antigen for diagnosis of bovine brucellosis.

    abstract::A radial immunodiffusion (RID) test employing a polysaccharide antigen (poly B) was compared with tests currently used in the diagnosis of bovine brucellosis. Over 1,000 sera from vaccinated and infected cattle, all of which had been examined bacteriologically, were used to determine the sensitivity and specificity of...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.12.6.753-760.1980

    authors: Jones LM,Berman DT,Moreno E,Deyoe BL,Gilsdorf MJ,Huber JD,Nicoletti P

    更新日期:1980-12-01 00:00:00

  • Predominance of the ac variant in K88-positive Escherichia coli isolates from swine.

    abstract::Monoclonal antibodies to K88ac and K88ab were used in enzyme-linked immunosorbent assays on Escherichia coli cultures known to produce K88 pili. A total of 415 K88-positive E. coli isolates from nine states were all found to be the K88ac variant. The cultures tested were isolated during the years 1976 to 1985. ...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.26.1.149-150.1988

    authors: Westerman RB,Mills KW,Phillips RM,Fortner GW,Greenwood JM

    更新日期:1988-01-01 00:00:00

  • Systematic investigation of enrichment media for wild-type Campylobacter jejuni strains.

    abstract::Of the media examined, thioglycolate broth supplemented with 5% lysed sheep blood, Butzler antibiotic mixture, and 0.1% lauryl sulfate was the most sensitive enrichment medium for recovery of wild-type strains of Campylobacter jejuni from cecal contents of chickens and chicken livers. It allowed the retrieval of 1 CFU...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.20.1.77-80.1984

    authors: Barot MS,Bokkenheuser VD

    更新日期:1984-07-01 00:00:00

  • Rapid speciation of Haemophilus with the porphyrin production test versus the satellite test for X.

    abstract::The detection of porphyrins produced from omega-aminolevulinic acid was evaluated for use in speciation of the Haemophilus. Two hundred and forty-eight clinical isolates of nonhemolytic Haemophilus were tested concurrently for porphyrin production and for X and V requirements. The porphyrin production test was more ra...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:

    authors: Lund ME,Blazevic DJ

    更新日期:1977-02-01 00:00:00

  • Extensive DNA sequence conservation throughout the Mycobacterium tuberculosis complex.

    abstract::The Mycobacterium tuberculosis complex includes the four species M. tuberculosis, M. bovis, M. africanum, and M. microti. We sequenced 13 M. tuberculosis complex strains in the 16S-to-23S rDNA internal transcribed spacer (ITS). The ITS has a high rate of nucleotide substitution. Previous reports found three nucleotide...

    journal_title:Journal of clinical microbiology

    pub_type: 杂志文章

    doi:10.1128/JCM.32.7.1639-1643.1994

    authors: Frothingham R,Hills HG,Wilson KH

    更新日期:1994-07-01 00:00:00