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Author De la Iglesia, R.; Valenzuela-Heredia, D.; Pavissich, J.P.; Freyhoffer, S.; Andrade, S.; Correa, J.A.; Gonzalez, B. pdf  doi
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  Title Novel polymerase chain reaction primers for the specific detection of bacterial copper P-type ATPases gene sequences in environmental isolates and metagenomic DNA Type
  Year 2010 Publication Letters In Applied Microbiology Abbreviated Journal Lett. Appl. Microbiol.  
  Volume 50 Issue 6 Pages 552-562  
  Keywords ATPases; bacterial communities; copper resistance; metagenomic DNA; PCR primer pair  
  Abstract Aims: In the last decades, the worldwide increase in copper wastes release by industrial activities like mining has driven environmental metal contents to toxic levels. For this reason, the study of the biological copper-resistance mechanisms in natural environments is important. Therefore, an appropriate molecular tool for the detection and tracking of copper-resistance genes was developed. Methods and Results: In this work, we designed a PCR primer pair to specifically detect copper P-type ATPases gene sequences. These PCR primers were tested in bacterial isolates and metagenomic DNA from intertidal marine environments impacted by copper pollution. As well, T-RFLP fingerprinting of these gene sequences was used to compare the genetic composition of such genes in microbial communities, in normal and copper-polluted coastal environments. New copper P-type ATPases gene sequences were found, and a high degree of change in the genetic composition because of copper exposure was also determined. Conclusions: This PCR based method is useful to track bacterial copper-resistance gene sequences in the environment. Significance and Impact of the Study: This study is the first to report the design and use of a PCR primer pair as a molecular marker to track bacterial copper-resistance determinants, providing an excellent tool for long-term analysis of environmental communities exposed to metal pollution.  
  Address [Gonzalez, B.] Univ Adolfo Ibanez, Fac Ingn & Ciencia, Santiago 7941169, Chile, Email: bernardo.gonzalez@uai.cl  
  Corporate Author Thesis  
  Publisher Wiley-Blackwell Publishing, Inc Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 0266-8254 ISBN Medium  
  Area Expedition Conference  
  Notes WOS:000277417000002 Approved  
  Call Number UAI @ eduardo.moreno @ Serial 88  
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Author Martinez, J.; Simon, V.; Gonzalez, B.; Conget, P. pdf  doi
openurl 
  Title A real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood Type
  Year 2010 Publication Letters In Applied Microbiology Abbreviated Journal Lett. Appl. Microbiol.  
  Volume 50 Issue 6 Pages 603-610  
  Keywords American foulbrood; flow cytometry; Paenibacillus larvae; real-time PCR; 16S rRNA specific primers  
  Abstract Aim: To develop a real-time PCR-based strategy for the detection of Paenibacillus larvae vegetative cells and spores to improve the diagnosis and the screening of American foulbrood (AFB), the most harmful pathology of honeybee brood. Methods and Results: A real-time PCR that allowed selective identification and quantification of P. larvae 16S rRNA sequence was developed. Using standard samples quantified by flow cytometry, detection limits of 37 center dot 5 vegetative cells ml-1 and 10 spores ml-1 were determined. Compared to spread plate method, this real-time PCR-based strategy allowed, in only 2 h, the detection of P. larvae in contaminated honeys. No false-positive results were obtained. Moreover, its detection limit was 100 times lower than that of the culture method (2 vs 200 spores g-1 of honey). Conclusion: A rapid, selective, with low detection limit, sensitive and specific method to detect and quantify vegetative cells and spores of P. larvae is now available. Significance and Impact of Study: In addition to honey samples, this real-time PCR-based strategy may be also applied to confirm AFB diagnosis in honeybee brood and to screen other apiary supplies and products (bees, pollen, wax), thus broadening the control of AFB spreading.  
  Address [Martinez, J.; Simon, V.; Conget, P.] Clin Alemana Univ Desarrollo, Fac Med, Inst Ciencias, Santiago 7710162, Chile, Email: pconget@udd.cl  
  Corporate Author Thesis  
  Publisher Wiley-Blackwell Publishing, Inc Place of Publication Editor  
  Language English Summary Language Original Title  
  Series Editor Series Title Abbreviated Series Title  
  Series Volume Series Issue Edition  
  ISSN 0266-8254 ISBN Medium  
  Area Expedition Conference  
  Notes WOS:000277417000009 Approved  
  Call Number UAI @ eduardo.moreno @ Serial 89  
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