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A novel and efficient assay for identification and quantification of Acidithiobacillus ferrooxidans in bioleaching samples

  • Biotechnology Methods
  • Published:
Journal of Industrial Microbiology & Biotechnology

Abstract

Acidithiobacillus ferrooxidans is a Gram-negative, acidophilic, and chemolithotrophic bacterium that is active in bioleaching. The leaching efficacy is directly influenced by the biomass changes of this specie in bioleaching microbial community. In order to perform a simple and sensitive assay on A. ferrooxidans from mixed strains in this process, a novel assay was developed based on sandwich hybridization assay with the aid of S1 nuclease treatment and fluorescent labeling. In the work, a designed DNA probe complementary to the conservative region of its 16S rRNA was synthesized, which showed high accuracy for distinguishing homologous species with the exclusion of even-only two base pairs difference. The specificity of this assay was verified in different systems with mixed strains, and the quantitative result was proved by comparison of microscopic cell counting. The detection sensitivity was about 8 × 102 cells/ml and the inter-assay coefficient of variation of three independent assays was from 3.8 to 7.7 %, respectively. In addition, the cycle of assay was about 3–4 h when the cost estimated was less than $0.5 per sample. This assay method might be applied for identifying and monitoring any kind of bacterial strain from a mixed microbial flora in bioleaching or other areas.

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Acknowledgments

This work was supported by grants from the National High Technology Research and Development Program of China (863 Program) (No. 2012AA021201), the Program of Innovation Projects Plan of Jiangsu Province (No. CXZZ11_0481), the Scientific Research Foundation for Graduate Students of Jiangnan University (No. JUDCF11013), the Program of the Science and Technology Support Plan of Jiangsu Province (No. SBE201077545 and SBE201170578), the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, the 111 Project (No. 111-2-06), the Scientific Research Foundation for Young Scholars of Jiangnan University (No. 2008LQN017), the Scientific Program of Jiangnan University (No. JUSRP11120 and JUSRP111A25), and the Program of the Key Laboratory of Industrial Biotechnology, Ministry of Education, China (No. KLIB-KF200906 and KLIB-KF 201006).

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Authors and Affiliations

  1. The Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, 1800 Lihu Road, Wuxi, 214122, People’s Republic of China

    Shoushuai Feng, Yu Xin, Hailin Yang, Ling Zhang, Wenliang Kang, Xiaole Xia & Wu Wang

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  1. Shoushuai Feng
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  2. Yu Xin
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  3. Hailin Yang
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  4. Ling Zhang
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  5. Wenliang Kang
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  6. Xiaole Xia
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  7. Wu Wang
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Correspondence to Wu Wang.

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Feng, S., Xin, Y., Yang, H. et al. A novel and efficient assay for identification and quantification of Acidithiobacillus ferrooxidans in bioleaching samples. J Ind Microbiol Biotechnol 39, 1161–1168 (2012). https://doi.org/10.1007/s10295-012-1118-9

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  • DOI: https://doi.org/10.1007/s10295-012-1118-9

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