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1.
Nicola Pozzato Elisabetta Stefani Katia Capello Riccardo Muliari Gaddo Vicenzoni 《World journal of microbiology & biotechnology》2011,27(1):31-37
Molecular diagnostic tests are widely implemented in animal and human health microbiology. Efficient nucleic acid extraction
methods are essential in diagnostic laboratories and automation is a valuable tool for those with high throughput activity.
Nucleic acid extraction protocols present variable efficiency depending on the composition of the specimen and the chemical-physical
characteristics of the target pathogen. In the present study, we compared the DNA extraction performances of four automated
methods (kits I, M, P, Q) adapted on a Hamilton Robotics “Microlab Starlet” extraction unit, and one manual method (kit R).
Ten-fold dilutions of Mycobacterium avium subsp. paratuberculosis (MAP) were used to contaminate bovine central nervous system (CNS) and lung-spleen-liver pools (LSL). In consideration of
its chemical-physical characteristics, MAP was selected as a template for pathogen DNA detection. Analytical performance and
repeatability were assessed through downstream real-time PCR amplification, hands-on time (HOT), total turnaround time (TAT)
and costs of all kits. MAP was detected differently depending on extraction kit and type of matrix analysed. Kits M and I
showed the highest analytical performance on CNS (1 MAP/ml) and LSL (10 MAP/ml), respectively. Besides analytical results,
kits I and M displayed high repeatability, the same HOT, very similar TAT, and were inexpensive. In conclusions, different
standardized automated systems have been established with high throughput, sensitivity and repeatability for CNS and LSL.
Our results also demonstrated that is necessary to assess the effectiveness of extraction kits in matrices not previously
tested to avoid the risk of unreliable diagnostic outcomes. 相似文献
2.
Amr El-Sayed Abdulwahed Ahmed Hassan Saleh Natour Amir Abdulmawjood Michael Bülte Wilfried Wolter Michael Zschöck 《Journal of microbiology (Seoul, Korea)》2009,47(3):253-259
The aim of the present study is to evaluate the efficiency of three methods to determine the molecular diversity of 34 Mycobacterium avium subsp. paratuberculosis (MAP) strains isolated from 17 cattle herds. The applied methods included the analysis of sequence polymorphism of the mononucleotide
(G1 and G2) and trinucleotide sequences (GGT) of the Short Sequence Repeats (SSR) and the determination of size polymorphism
of 9 different Mycobacterial Interspersed Repetitive Units (MIRU) and 6 Variable Number Tandem Repeats (VNTR). Sequence analysis
of SSR of 34 isolates showed 4, 6, and 2 alleles of G1, G2, and GGT repeats, respectively. The amplification of the investigated
9 MIRU units revealed only two discriminatory genotyping systems (MIRU2 and MIRU3). Out of 6 VNTR PCR differentiation methods,
only one method could be recommended for genotyping purposes. The profile 7g-12g-4ggt-II-b-2 of the combination systems G1-G2-GGT-MIRU2-MIRU3-VNTR1658
dominates among the examined isolates and was detected in 14.7% of the isolates. The use of certain repetitive loci of SSR,
MIRU, and VNTR techniques in this study showed greater potential than others for the characterization of MAP isolates. The
recommended loci can be used for the epidemiological tracing of MAP field strains and to determine the relationships between
isolates in different herds. 相似文献
3.
Quantitative trait loci (QTL) detection was carried out for adventitious rooting and associated propagation traits in a second-generation
outbred Corymbia torelliana × Corymbia citriodora subspecies variegata hybrid family (n = 186). The parental species of this cross are divergent in their capacity to develop roots adventitiously on stem cuttings
and their propensity to form lignotubers. For the ten traits studied, there was one or two QTL detected, with some QTL explaining
large amounts of phenotypic variation (e.g. 66% for one QTL for percentage rooting), suggesting that major effects influence
rooting in this cross. Collocation of QTL for many strongly genetically correlated rooting traits to a single region on linkage
group 12 suggested pleiotropy. A three locus model was most parsimonious for linkage group 12, however, as differences in
QTL position and lower genetic correlations suggested separate loci for each of the traits of shoot production and root initiation.
Species differences were thought to be the major source of phenotypic variation for some rooting rate and root quality traits
because of the major QTL effects and up to 59-fold larger homospecific deviations (attributed to species differences) relative
to heterospecific deviations (attributed to standing variation within species) evident at some QTL for these traits. A large
homospecific/heterospecific ratio at major QTL suggested that the gene action evident in one cross may be indicative of gene
action more broadly in hybrids between these species for some traits. 相似文献
4.
There are three most important bacterial causative agents of serious infections that could be misused for warfare purposes:
Bacillus anthracis (the causative agent of anthrax) is the most frequently mentioned one; however, Fracisella tularensis (causing tularemia) and Yersinia pestis (the causative agent of plague) are further bacterial agents enlisted by Centers for Disease Control and Prevention into the category A of potential biological weapons. This review intends to summarize basic information about these bacterial
agents. Military aspects of their pathogenesis and the detection techniques suitable for field use are discussed. 相似文献
5.
N. Yu. Markelova 《Microbiology》2010,79(6):777-779
Interaction of Bdellovibrio bacteriovorus 100NCJB with bacteria Campylobacter jejuni (strains 1, 2, 3, 4, and 5) and Helicobacter pylori, strain TX30a, was confirmed. The results indicate that lytic activity of bdellovibrios both in liquid media and cells attached
to a surface was observed. The potential use of the antimicrobial activity of predatory bacteria for environmental bioprotection
and public health is discussed. 相似文献
6.
O. V. Golovanova V. I. Konenkov A. V. Shevchenko M. V. Smolnikova 《Russian Journal of Genetics》2009,45(8):981-986
Based on population analysis of the DRB1, DQA1, DQB1 and TNFA allele frequency distribution patterns, regional features of immunogenetic structure of the population of West Siberia were
investigated. Statistically significant linkage disequilibrium within the HLA class II region, as well as between the TNFA and DRB1, DQA1, and DQB1 was demonstrated. Population frequency distribution patterns of two- and multilocus haplotypes were examined. 相似文献
7.
8.
To overexpress the chitosanase gene (csn) in F. solani, a vector based on pCAMBIA 1300 was constructed. The csn gene, which is under control of the Aspergillus nidulans
gpdA promoter and A. nidulans trpC terminator, was introduced back into the F. solani genome by Agrobacterium tumefaciens-mediated transformation, and the herbicide-resistance gene bar from Streptomyces hygroscopicus was used as the selection marker. Transformants which showed a significant increase in chitosanase production (~2.1-fold
than control) were obtained. Southern blot analysis indicated that most transformants had a single-copy T-DNA integration. 相似文献
9.
Hiroyoshi Kubo 《Mycoscience》2009,50(5):400-406
Pilobolus crystallinus shows unique photoresponses at various growing stages. cDNAs for putative photoreceptors were cloned from this fungus. Three
genes named pcmada1, pcmada2, and pcmada3 were identified from the PCR fragments, and amplified with degenerated primers for the LOV domain, which is conserved in
many blue-light receptors. Deduced amino acid sequences for PCMADA1, PCMADA2, and PCMADA3 had one light-oxygen-voltage (LOV)-sensing
and two PER-ARNT-SIM (PAS) domains. A zinc finger DNA-binding motif was conserved in the C-terminals of PCMADA1 and PCMADA3.
However, PCMADA2 lacked the zinc finger motif. Expression of pcmada1 was suppressed by blue light whereas that of pcmada3 was promoted by blue-light irradiation. 相似文献
10.
The intron sequence of chloroplast rpS16 and the secondary structure of its pre-mRNA were characterized for the first time in 26 Allium sativum accessions of different ecologo-geographical origins and seven related Allium species. The boundaries and main stem-loop consensus sequences were identified for all six domains of the intron. Polymorphism
was estimated for the total intron and its regions. The structural regions of the rpS16 intron proved to be heterogeneous for mutation rate and spectrum. Mutations were most abundant in domains II and IV, and
transition predominated in domains I, III, V, and VI. In addition to structural elements and motifs typical for group IIB
introns, several Allium-specific micro- and macrostructural mutations were revealed. A 290-bp deletion involving domains III and IV and part of domain
V was observed in A. altaicum, A. fistulosum, and A. schoenoprasum. Several indels and nucleotide substitutions were found to cause a deviation of the pre-mRNA secondary structure from the
consensus model of group II introns. 相似文献
11.
12.
I. Rosario G. Soro S. Déniz O. Ferrer F. Acosta D. Padilla B. Acosta 《Mycopathologia》2010,169(4):315-319
Columba livia is an important reservoir and carrier of Cryptococcus neoformans, Cryptococcus uniguttulatus, Cryptococcus laurentii and Cryptococcus albidus. Upper digestive tract of this species is also known as a habitat for Cryptococcus neoformans. Given the increasing clinical interest of this microorganism, 331 swabs from crop and 174 dropping samples from pigeon lofts
in Grand Canary Island have been studied. The obtained results show an extensive presence samples 81 positive (24.47%) of
Cryptococcus spp. in analysed crops: 32 (9.66%) for C. neoformans, 24 (7.2%) for C. uniguttulatus, 23 (6.9%) for C. albidus and 2 (0.6%) for C. laurentii. In the same way, Cryptococcus spp was also isolated in 82 (47.13%), dropping samples: C. neoformans in 59 (33.9%), C. uniguttulatus, in 9 (5.17%), C. laurentii in 8 (4.59%) and C. albidus in 6 (3.44%) of the investigated samples, respectively. The cryptococcosis produced by species of cryptococci other than C. neoformans has become more important during the last decade, supporting the study on the role of pigeon in the epidemiology of this
disease. 相似文献
13.
Nanobodies (or VHHs) are single-domain antigen-binding fragments derived from Camelid heavy chain-only antibodies. Their small size, monomeric behaviour, high stability and solubility, and ability to bind epitopes
not accessible to conventional antibodies make them especially suitable for many therapeutic and biotechnological applications.
Here we describe high-level expression, in Nicotiana benthamiana, of three versions of an anti-hen egg white lysozyme (HEWL) nanobody which include the original VHH from an immunized library
(cAbLys3), a codon-optimized derivative, and a codon-optimized hybrid nanobody comprising the CDRs of cAbLys3 grafted onto
an alternative ‘universal’ nanobody framework. His6- and StrepII-tagged derivatives of each nanobody were targeted for accumulation
in the cytoplasm, chloroplast and apoplast using different pre-sequences. When targeted to the apoplast, intact functional
nanobodies accumulated at an exceptionally high level (up to 30% total leaf protein), demonstrating the great potential of
plants as a nanobody production system. 相似文献
14.
Following a re-examination of the material treated under Barleria brevispina (Fiori) Hedrén in the recent Flora of Somalia account of the Acanthaceae, it is concluded that two distinct species are involved and Barleria compacta Malombe & I. Darbysh. is described here from north-eastern Somalia. Its affinities and conservation status are discussed. 相似文献
15.
Gaye Öngen Gaye Güngör Bahar Kanberoglu 《World journal of microbiology & biotechnology》2007,23(4):519-524
Aspergillus section Nigri strains Aspergillus aculeatus Ege-K 258, A.
foeditus var. pallidus Ege-K156, A. niger Ege-K 4 and A. tubingensis Ege-K 265 were used to treat olive mill wastewater (OMW) in an investigation aimed at exploring their dephenolisation and
decolourisation ability and, consequently, the economic feasibility of using any or all of these strains in a pre-treatment
step in the processing of OMW. Of these strains A. tubingensis Ege-K 265 resulted in an 80% decolourisation of twofold-diluted OMW and a 30% decolourisation of undiluted OMW; in addition,
it was able to remove approximately 30% of all phenolic compounds in both twofold-diluted and undiluted OMW. We conclude that
A. tubingensis Ege-K 265 could be effectively used in the pre-treatment step of a combined aerobic-anaerobic process to solve the environmental
problems caused by OMW in Mediterranean countries. 相似文献
16.
17.
Swasti S. Swain Tapasi Tripathy Pradipta K. Mohapatra Pradeep K. Chand 《In vitro cellular & developmental biology. Plant》2010,46(2):134-141
In vitro regeneration of black nightshade (Solanum nigrum L.) plants was achieved through callus-mediated shoot organogenesis followed by 30 d indoor ex vitro adaptation to nutritional stress under environmental ambience and thereafter 6-d outdoor acclimatization in pots prior to
field establishment. Relevant physiological parameters including pigment content, chlorophyll a fluorescence, net photosynthetic rate (P
N), transpiration rate (E), and stomatal conductance (g
s) of in vitro-regenerated plants were investigated during the course of ex vitro adaptation. During the first 4 d of indoor transplantation to potting substrate, there was a marginal reduction in the leaf
chlorophyll and carotenoid contents but P
N and E were strongly reduced. The stomatal conductance and E/P
N ratio were significantly higher in plants up to 20 d of indoor adaptation than those of comparable age grown naturally from
seeds. The shape of the OJIP fluorescence transient varied significantly with acclimatization, and the maximum change was
observed at 2.0 ms. The 2.0 ms variable fluorescence (V
j), 30 ms relative fluorescence (M
0), photon trapping probability (TR0/Abs), and photosystem II (PSII) trapping rate (TR0/RC) showed initial disturbance and subsequent stabilization during 30 d of indoor acclimatization. Energy dissipation (DI0/RC) and electron transport probability (ET0/TR0) showed an initial phase of increase during the 4 d after plants were transplanted outdoors. During the 6-d outdoor acclimatization
after transfer of plants to soil, no significant change in total chlorophylls and carotenoids, E, and g
s were observed, but P
N improved after reduction on the first d. The OJIP-derived parameters experienced change on the first d but were stabilized
quickly thereafter. There was no significant difference between outdoor acclimatized plants and those of the seed-grown plants
of comparable age with respect to photosynthetic and fluorescence parameters. Direct transfer of plants without indoor acclimatization,
however, showed a completely different trend with respect to P
N, E, and OJIP fluorescence transients. The bearing of this study on optimizing micropropagation is discussed. 相似文献
18.
19.
Ching-Nan Lin Wan-Jr Syu Wei-Sheng W Sun Jenn-Wei Chen Tai-Hung Chen Ming-Jaw Don Shao-Hung Wang 《Journal of biomedical science》2010,17(1):84
Plumbagin is found in many herbal plants and inhibits the growth of various bacteria. Escherichia coli strains are relatively resistant to this drug. The mechanism of resistance is not clear. Previous findings showed that plumbagin
treatment triggered up-regulation of many genes in E. coli including ahpC, mdaB, nfnB, nfo, sodA, yggX and ygfZ. By analyzing minimal inhibition concentration and inhibition zones of plumbagin in various gene-disruption mutants, ygfZ and sodA were found critical for the bacteria to resist plumbagin toxicity. We also found that the roles of YgfZ and SodA in detoxifying
plumbagin are independent of each other. This is because of the fact that ectopically expressed SodA reduced the superoxide
stress but not restore the resistance of bacteria when encountering plumbagin at the absence of ygfZ. On the other hand, an ectopically expressed YgfZ was unable to complement and failed to rescue the plumbagin resistance
when sodA was perturbed. Furthermore, mutagenesis analysis showed that residue Cys228 within YgfZ fingerprint region was critical for
the resistance of E. coli to plumbagin. By solvent extraction and HPLC analysis to follow the fate of the chemical, it was found that plumbagin vanished
apparently from the culture of YgfZ-expressing E. coli. A less toxic form, methylated plumbagin, which may represent one of the YgfZ-dependent metabolites, was found in the culture
supernatant of the wild type E. coli but not in the ΔygfZ mutant. Our results showed that the presence of ygfZ is not only critical for the E coli resistance to plumbagin but also facilitates the plumbagin degradation. 相似文献