Design of a species-specific PCR method for the detection of the heat-resistant fungi Talaromyces macrosporus
 and Talaromyces trachyspermus

The aim of this study was to report the detection of methicillin-resistant Staphylococcus aureus (MRSA) strains at a veterinary university goat breeding farm and their characteristics. A total of 278 samples collected from animals, milk, environment and farm personnel between June 2006 and March 2008 were examined. The identification of S. aureus isolates was performed by a species specific PCR assay. All detected isolates were tested for resistance to oxacillin and other antimicrobials by phenotypic methods and for the mecA gene by PCR method. Eight MRSA were detected in this study. Five of them originated from goat’s milk and three were recovered from one human carrier of the farm personnel. All obtained MRSA isolates were clonally consistent and were characterized as: SCCmec type IV, spa type t064, seb positive and for genes encoding TSST-1, PVL and exfoliative toxins A and B negative.

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Production of biogenic amines by Enterococci

Czech Journal of Food Sciences ◽

10.17221/673-cjfs ◽

2010 ◽

Vol 27 (Special Issue 2) ◽

pp. 50-55 ◽

Cited By ~ 4

Author(s):

K. Kučerová ◽

H. Svobodová ◽

Š. Tůma ◽

I. Ondráčková ◽

M. Plocková

Keyword(s):

Growth Medium ◽

Histidine Decarboxylase ◽

Cow Milk ◽

Decarboxylase Activity ◽

Differential Growth ◽

Specific Pcr ◽

Enterococcus Spp ◽

Pcr Method ◽

Species Specific ◽

Histidine Decarboxylase Activity

Enterococci were presented in all tested samples of raw cow milk (six samples) at the level 103–105 CFU/ml, fresh cheeses (five samples) at the level 102–106 CFU/g and semi-hard cheeses (five samples) at the level 103–105 CFU/g. All 33 isolated Enterococcus strains were screened for decarboxylase activity by usage differential growth medium and 20 of them possessed tyrosine decarboxylase activity. A collection of eight strains with the strongest decarboxylase activity were identified by species specific PCR as E. faecium (Z3, Z4, Br4 and 6/4D strains) and E. faecalis (Ž4, 3/3C and 4/1A strains). Enterococcus spp. Z1 strain was not identified at the species level by used methods, but the genus was confirmed by PCR method. Their tyrosin decarboxylase activity was confirmed by TLC and detection of tdc gene. Z1, Z3 and Z4 strains showed also histidine decarboxylase activity on the differential growth medium with histidine, but this activity was evaluated by TLC as a false positive reaction of medium.

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Design of species-specific PCR method for the detection of pathogenEmbellisia astragaliin standing milk vetch seeds

Letters in Applied Microbiology ◽

10.1111/lam.12381 ◽

2015 ◽

Vol 60 (4) ◽

pp. 372-378 ◽

Cited By ~ 1

Author(s):

J.L. Liu ◽

Y.Z. Li ◽

Z.B. Nan

Keyword(s):

Specific Pcr ◽

Pcr Method ◽

Species Specific

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Species-specific PCR method for identification of Streptococcus downei

Letters in Applied Microbiology ◽

10.1111/j.1472-765x.2003.01451.x ◽

2004 ◽

Vol 38 (2) ◽

pp. 125-129 ◽

Cited By ~ 2

Author(s):

T. Igarashi ◽

E. Asaga ◽

C. Murai ◽

R. Sasa ◽

N. Goto

Keyword(s):

Specific Pcr ◽

Pcr Method ◽

Species Specific

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Detection of Phomopsis sclerotioides in Commercial Cucurbit Field Soil by Nested Time-Release PCR

Plant Disease ◽

10.1094/pdis-03-11-0187 ◽

2012 ◽

Vol 96 (4) ◽

pp. 515-521 ◽

Cited By ~ 3

Author(s):

T. Ito ◽

S. Fuji ◽

E. Sato ◽

Y. Iwadate ◽

T. Toda ◽

...

Keyword(s):

Minimum Concentration ◽

Disease Symptoms ◽

Specific Pcr ◽

Phomopsis Sclerotioides ◽

Fluorescent Pcr ◽

Pcr Products ◽

Pcr Method ◽

Polymerase Chain ◽

Species Specific ◽

Pcr Techniques

A polymerase chain reaction (PCR)-based molecular method to detect Phomopsis sclerotioides in soil was developed using a species-specific primer pair. To improve sensitivity of the detection, three PCR techniques were used; namely, nested PCR using the primer pair internal transcribed spacer (ITS)1 and ITS4, time-release PCR using two different DNA polymerases (recombinant Taq and AmpliTaq Gold), and fluorescent PCR to obtain fluorescent-labeled PCR products that can be analyzed by capillary electrophoresis. The latter two techniques were combined and termed nested time-release fluorescent (NTRF)-PCR. The minimum concentration of DNA required to obtain species-specific PCR products successfully was 50 fg/μg. Using the NTRF-PCR method, the fungus could be detected in sandy soil that was artificially infested at a density of 10 CFU/g. The pathogen was detected in most soil samples collected from commercial cucumber fields in which visual disease symptoms had appeared, and even in samples collected from fields where visual disease symptoms had not appeared. To prevent the invasion and establishment of root-inhabiting pathogens such as P. sclerotioides, it is critical to detect the fungus in soil as soon as possible after its introduction into a cucumber-growing region.

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