Abstract:
:To date, no single reported DNA extraction method is suitable for the efficient extraction of DNA from all fungal species. The efficiency of extraction is of particular importance in PCR-based medical diagnostic applications where the quantity of fungus in a tissue biopsy may be limited. We subjected 16 medically relevant fungi to physical, chemical and enzymatic cell wall disruption methods which constitutes the first step in extracting DNA. Examination by light microscopy showed that grinding with mortar and pestle was the most efficient means of disrupting the rigid fungal cell walls of hyphae and conidia. We then trialled several published DNA isolation protocols to ascertain the most efficient method of extraction. Optimal extraction was achieved by incorporating a lyticase and proteinase K enzymatic digestion step and adapting a DNA extraction procedure from a commercial kit (MO BIO) to generate high yields of high quality DNA from all 16 species. DNA quality was confirmed by the successful PCR amplification of the conserved region of the fungal 18S small-subunit rRNA multicopy gene.
journal_name
J Microbiol Methodsjournal_title
Journal of microbiological methodsauthors
Karakousis A,Tan L,Ellis D,Alexiou H,Wormald PJdoi
10.1016/j.mimet.2005.06.008subject
Has Abstractpub_date
2006-04-01 00:00:00pages
38-48issue
1eissn
0167-7012issn
1872-8359pii
S0167-7012(05)00183-1journal_volume
65pub_type
杂志文章abstract::The level of microbial resistance to heavy metals is an important issue for the microbial ecology of heavy metal-contaminated habitats. However, assays based upon growth in nutrient media will overestimate the resistance level due to metal ion interactions with inorganic and organic components. The analysis of Pb-resi...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(99)00032-9
更新日期:1999-07-01 00:00:00
abstract::Bacterial polyhydroxyalkanoate (PHA) is an attractive biopolyester for medical applications due to its biocompatibility. However, inappropriate extraction of PHA from bacterial biomass results in contamination by pyrogenic compounds (e.g. lipopolysaccharides) and thus influences medical testing. This problem was solve...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2007.01.002
更新日期:2007-04-01 00:00:00
abstract::Due to technical problems, biofilm biomasses are difficult to be precisely determined. One reliable strategy is based on the colorimetry of formazan compounds derived from tetrazolium salt reduction. XTT presents some desirable properties that make the biofilm measurements easier. However, cells entrapped within the e...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2010.12.018
更新日期:2011-02-01 00:00:00
abstract::Cyclospora cayetanensis, a coccidian parasite, with a fecal-oral life cycle, has become recognized worldwide as an emerging human pathogen. Clinical manifestations include prolonged gastroenteritis. While most cases of infection with C. cayetanensis in the United States have been associated with foodborne transmission...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(02)00209-9
更新日期:2003-04-01 00:00:00
abstract::A rapid and simple DNA labeling system has been developed for disposable microarrays and has been validated for the detection of 117 antibiotic resistance genes abundant in Gram-positive bacteria. The DNA was fragmented and amplified using phi-29 polymerase and random primers with linkers. Labeling and further amplifi...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.11.006
更新日期:2015-01-01 00:00:00
abstract::We devised a microbial culture apparatus capable of maintaining sub-nanomolar H2 concentrations. This apparatus provides a method for study of interspecies hydrogen transfer by externally fulfilling the thermodynamic requirement for low H2 concentrations, thereby obviating the need for use of cocultures to study some ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(99)00125-6
更新日期:2000-02-01 00:00:00
abstract::RNA-based molecular technique (RT-qPCR) is a promising method for microcystin monitoring in lakes and reservoirs, but great lability of RNA in cyanobacterial samples limits its application. To date, no studies have investigated how to effectively preserve RNA in cyanobacterial samples. In this study, four different tr...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2019.105684
更新日期:2019-09-01 00:00:00
abstract::Several outbreaks of foodborne yersiniosis have been documented and this disease continues to be source of infections transmitted through foods. The selective agars most commonly used to isolate Yersinia enterocolitica in clinical, food and environmental samples, cefsulodin-irgasan-novobiocin (CIN) and MacConkey (MAC)...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2007.11.019
更新日期:2008-02-01 00:00:00
abstract::Molecular typing is an important tool in surveillance and outbreak investigations of human Salmonella infections. In this study, three molecular typing methods were used to investigate the discriminatory ability, reproducibility and the genetic relationship between 110 Salmonella enterica subspecies enterica isolates....
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.03.006
更新日期:2005-11-01 00:00:00
abstract::In recent years, the rapid advances of culture-independent methods and new molecular tools have revolutionized our understanding of microbial biodiversity and ecological functions. DNA extraction from microbial communities is a critical step in this process and several methods have been proposed and used, but the infl...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2019.02.005
更新日期:2019-04-01 00:00:00
abstract::The development of biofilms in static plates was monitored. Glass coupons were placed on agar covered with filter paper, which was inoculated with suspended bacteria. The viable cell density, biofilms matrix and biomass were quantified. The method is excellent for adhesion and material studies, due to its simplicity a...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.10.008
更新日期:2014-12-01 00:00:00
abstract::Time of Flight-Secondary Ion Mass Spectrometry (ToF-SIMS) was used to study compositional characteristics of Extracellular Polymeric Substances (EPS) and compared these to characteristics of the EPS-matrix of intact diatom biofilms. Three benthic diatoms species were investigated, Cylindrotheca closterium, Navicula mu...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.10.001
更新日期:2006-06-01 00:00:00
abstract::We have developed a method to identify and amplify differential fragments between two complex genomes. This technique, named hybridization-monitored genome differential analysis (HMDA), incorporates a monitor system into a PCR-based solid subtraction hybridization that tracks the entire hybridization process. This is ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.05.010
更新日期:2006-03-01 00:00:00
abstract::In recent years the global rise in antibiotic resistance and environmental consciousness lead to a renewed fervour to find and develop novel antibiotics, including antifungals. However, the influence of the environment on antifungal activity is often disregarded and many in vitro assays may cause the activity of certa...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2012.09.025
更新日期:2012-12-01 00:00:00
abstract::To establish the gene transfer system in the cyanobacteria, we successfully introduced the shuttle plasmid pRL489 into Oscillatoria MKU 277 by electroporation with transformation frequencies of up to 102 cfu/mug of plasmid DNA. These findings suggest that Oscillatoria MKU 277 can be used as an experimental tool for ge...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2005.11.011
更新日期:2006-07-01 00:00:00
abstract::5'-Nuclease and a hybridization probe assays for the detection of shiga toxin-producing Escherichia coli were validated with regard to selectivity, analytical sensitivity, reproducibility and clinical performance. Both assays were capable of detecting the classical stx(1) and stx(2) genes when challenged with referenc...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2007.05.016
更新日期:2007-09-01 00:00:00
abstract::Using two-step-PCR screening which consists of direct and priority-based PCR and inverse PCR, fungal gene-deletion mutants were selected quickly and accurately. It omits genomic DNA extraction and Southern blotting steps and prevents misinterpretations caused by PCR failure. It is anticipated to facilitate large-scale...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.06.023
更新日期:2014-10-01 00:00:00
abstract::Tropheryma whipplei is the causative agent of Whipple's disease (WD), a chronic, life-threatening infection. Laboratory diagnosis is mainly based on PCR and histopathological analysis in duodenal biopsies and other specimens requiring invasive procedures. We have examined the presence of antibodies to recombinant heat...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/s0167-7012(01)00336-0
更新日期:2001-12-01 00:00:00
abstract::The paper and pulp industry (PPI) produces high quantities of solid and liquid discharge and is regarded as the most polluting industry in the world causing adverse effects to environments and human beings. Hence changes in the way PPI sludge and waste materials are treated is urgently required. Nearly, 10 million ton...
journal_title:Journal of microbiological methods
pub_type: 杂志文章,评审
doi:10.1016/j.mimet.2019.03.015
更新日期:2019-05-01 00:00:00
abstract::Rapid identification of bacterial pathogens facilitates earlier optimization of antibiotic treatment and reduces morbidity and mortality in sepsis patients. The aim of this research was to design an in-house chemical-free method for direct bacterial identification in positive blood culture (BC) broths and to compare t...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2020.106065
更新日期:2021-01-01 00:00:00
abstract::A strain-specific assay was developed for the detection of viable Lactobacillus on cattle feed. The DNA sequences of the 16S rRNA gene and four different 16S/23S rRNA intergenic spacer regions (ISR) from Lactobacillus sp. HOFG1 were determined. Based on these sequences, a strain-specific primer was designed for the am...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2004.12.002
更新日期:2005-05-01 00:00:00
abstract::A new, simple approach for the isolation and quantitative analysis of the cell wall (CW) proteins from invasively growing Saccharomyces cerevisiae strains is described in this contribution. The proposed method was proved compatible with agar-invasion assays and was demonstrated to be useful as a screening tool for rap...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2009.09.003
更新日期:2009-12-01 00:00:00
abstract::There is a paucity of knowledge on gene expression in dead bacterial cells. Why would this knowledge be useful? The cells are dead. However, the time duration of gene expression following cell death is often unknown, and possibly in the order of minutes. In addition, it is a challenge to determine if bacterial cells a...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2012.04.004
更新日期:2012-07-01 00:00:00
abstract::The aim of this study was to (i) design, develop and validate a practical and physiologically relevant reconstituted in vitro oral mucosa tissue model and (ii) to assess its applicability in in vitro host-pathogen interactions with C. albicans and S. aureus. Co-culture organotypic constructions were created by incorpo...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2018.07.004
更新日期:2018-09-01 00:00:00
abstract::The quality and quantity of sputum collected has an important impact on the laboratory diagnosis of pulmonary TB. We conducted a pilot study to assess a new collection cups for the collection of sputum for the diagnosis of pulmonary tuberculosis. The pilot study utilized the standard collection cup in South Africa dem...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2017.01.011
更新日期:2017-04-01 00:00:00
abstract::Traditional antibiotic susceptibility testing methods take several days to confirm and start disease treatment. The lack of new antibiotics or drugs warrants a need for optimization of current diagnostic tools for immediate antibiotic susceptibility testing. We present a rapid screening method to evaluate the response...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2019.05.009
更新日期:2019-07-01 00:00:00
abstract::MALDI-TOF mass spectrometry (MALDI-TOF MS) is increasingly used as a reliable technique for species identification of bacterial pathogens. In this study we investigated the question of whether MALDI-TOF MS can be used for accurate sub-differentiation of strains and isolates of two important nosocomial pathogens Entero...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2014.02.015
更新日期:2014-05-01 00:00:00
abstract::Host DNA may adversely affect metagenomic studies focusing on the prokaryotic microbiota. This study compared the levels of host DNA in subgingival plaque collected by paper points and curette, using quantitative PCR. Lower proportions of host DNA and higher proportions of bacterial DNA were recovered from samples col...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2015.01.023
更新日期:2015-04-01 00:00:00
abstract::Staphylococcus aureus remains a leading, virulent pathogen capable of expressing complex drug resistance that requires up to 2-4 days for laboratory analysis. In this study, we evaluate the ability of automated microscopy of immobilized live bacterial cells to differentiate susceptible from non-susceptible responses o...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2013.12.021
更新日期:2014-03-01 00:00:00
abstract::The antimicrobial activity of lime oil was significantly enhanced against Staphylococcus epidermidis in a pulsed electromagnetic field. These findings support other workers who found that electromagnetic fields augment antimicrobial activity. ...
journal_title:Journal of microbiological methods
pub_type: 杂志文章
doi:10.1016/j.mimet.2010.09.015
更新日期:2010-11-01 00:00:00