scholarly journals Development of a Real-Time qPCR Method for Detection and Enumeration of Mycobacterium spp. in Surface Water

2010 ◽  
Vol 76 (21) ◽  
pp. 7348-7351 ◽  
Author(s):  
Nicolas Radomski ◽  
Françoise S. Lucas ◽  
Régis Moilleron ◽  
Emmanuelle Cambau ◽  
Sophie Haenn ◽  
...  
Keyword(s):  
Surface Water ◽  
16S Rrna ◽  
Real Time ◽  
Quantitative Pcr ◽  
Environmental Samples ◽  
Real Time Quantitative Pcr ◽  
Qpcr Method ◽  
Pcr Method ◽  
Real Time Qpcr ◽  
Mycobacterium Spp

ABSTRACT A real-time quantitative PCR method was developed for the detection and enumeration of Mycobacterium spp. from environmental samples and was compared to two other methods already described. The results showed that our method, targeting 16S rRNA, was more specific than the two previously published real-time quantitative PCR methods targeting another 16S rRNA locus and the hsp65 gene (100% versus 44% and 91%, respectively).

scholarly journals DETECTION, TRANSMISSION, AND EXPRESSION cDNA GROWTH HORMONE GENE (PhGH) OF STRIPPED CATFISH IN F-1 TRANSGENIC AFRICAN CATFISH

2014 ◽  
Vol 9 (2) ◽  
pp. 89
Author(s):  
Huria Marnis ◽  
Bambang Iswanto ◽  
Rommy Suprapto ◽  
Imron Imron ◽  
Raden Roro Sri Pudji Sinarni Dewi
Keyword(s):  
Growth Hormone ◽  
Real Time ◽  
Quantitative Pcr ◽  
African Catfish ◽  
Growth Hormone Gene ◽  
Real Time Quantitative Pcr ◽  
Qpcr Method ◽  
Gh Gene ◽  
Pcr Method ◽  
Transgenic Male

In previous study, the fast growth transgenic founder of African catfish was produced harboring a growth hormone (GH) gene construct containing a stripped catfish growth hormone (PhGH) cDNA. This study was conducted to investigate transgene (PhGH) transmission and expression in F-1 transgenic African catfish. The transgenic founders (female) were crossed with non-transgenic (male) to produce heterozygous F-1 progeny. PhGH gene was detected in the embryo, larvae, and seed of the transgenic F-1 using PCR method. Expression levels of transgene in embryo and larvae were analyzed using real-time quantitative PCR (qPCR) method. The transgene was detected in embryo, larvae and seed of F-1 transgenic African catfish. Founder could transmit PhGH gene to transgenic F-1 lines in ranged 36% to 48%. Expression level of Phgh gene in embryo was higher than that of the larvae; whereas in the embryo was 1.5 x 105 - 5.2 x 105 copies or 0.49-9.82 fold, while in the larvae was 1.1 x 105 - 2.5 x 105copies or 0.19-5.80 fold. 

Real-Time Quantitative PCR Detection of Mycobacterium avium Subspecies in Meat Products

2011 ◽  
Vol 74 (4) ◽  
pp. 636-640 ◽  
Author(s):  
B. KLANICOVA ◽  
I. SLANA ◽  
H. VONDRUSKOVA ◽  
M. KAEVSKA ◽  
I. PAVLIK
Keyword(s):  
Real Time ◽  
Quantitative Pcr ◽  
Mycobacterium Avium ◽  
General Public ◽  
Meat Products ◽  
Mycobacterium Avium Subsp ◽  
Insertion Sequences ◽  
Real Time Quantitative Pcr ◽  
Qpcr Method ◽  
Meat Samples

The aim of this work was to examine various purchased meat products and to find out if any traces of Mycobacterium avium subsp. avium, M. avium subsp. hominissuis, and M. avium subsp. paratuberculosis could be detected in these samples. Analysis of the meat products (raw, cooked, and fermented) was performed using a real-time quantitative PCR (qPCR) method for the detection of specific insertion sequences: duplex qPCR for the detection of IS900 specific for M. avium subsp. paratuberculosis, and triplex qPCR for the detection of IS901 specific for Mycobacterium avium subsp. avium and IS1245 specific for M. avium subsp. hominissuis. Of the 77 analyzed meat samples, 17 (22%) were found to contain M. avium subsp. paratuberculosis DNA, 4 (5%) samples contained Mycobacterium avium subsp. avium DNA, and in 12 (16%) samples M. avium subsp. hominissuis DNA was detected. The concentration of M. avium subsp. paratuberculosis and M. avium subsp. hominissuis DNA in some meat products exceeded 104 genomes per g. Culture examination of these mycobacterial subspecies was negative. By analyzing a range of meat products, we have provided evidence to support the hypothesis that M. avium is present in everyday commodities sold to the general public

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