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Sequence-specific electrochemical detection of double-strand PCR amplicons of PML/RARα fusion gene in acute promyelocytic leukemia

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Abstract

A novel electrochemical method for the sequence-specific detection of double-stranded polymerase chain reaction (PCR) products of PML/RARα fusion gene in acute promyelocytic leukemia (APL) was described in detail. Based on a “sandwich” sensing mode involving a pair of locked nucleic acids probes (capture probe and reporter probe), this DNA sensor exhibited excellent selectivity and specificity. The direct and quantitative analysis of double-stranded complementary was firstly performed by our sensor without the use of alkali, helicase enzymes, or denaturants. Finally, combining PCR technique with electrochemical detection scheme, PCR amplicons (191 bp) of the PML/RARα fusion gene were obtained and rapidly identified with a low detection limit of 79 fmol in the 100-μL hybridization system. The results clearly showed the power of sensor as a promising tool for the sensitive, specific, and portable detection of APL and other diseases.

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References

  1. de The H, Lavau C, Marchio A, Chomienne C, Degos L, Dejean A (1991) The PML-RARα fusion mRNA generated by the t(15;17) translocation in acute promyelocytic leukemia encodes a functionally altered RAR. Cell 66:675–684

    Article  Google Scholar 

  2. Choughule A, Polampalli S, Amre P, Shinde S, Banavali S, Prabhash K, Nair R, Subramanian PG, Gujral S, Parikh PM (2009) Identification of PML/RAR alpha fusion gene transcripts that showed no t(15;17) with conventional karyotyping and fluorescent in situ hybridization. Genet Mol Res 8:1–7

    Article  CAS  Google Scholar 

  3. Han JY, Kim KE, Kim KH, Park JI, Kim JS (2007) Identification of PML–RARA rearrangement by RT-PCR and sequencing in an acute promyelocytic leukemia without t(15;17) on G-banding and FISH. Leuk Res 31:239–243

    Article  CAS  Google Scholar 

  4. Wong ML, Medrano JF (2005) Real-time PCR for MRNA quantitation. Biotechniques 39:75–85

    Article  CAS  Google Scholar 

  5. Yogeswaran U, Chen SM (2008) A review on the electrochemical sensors and biosensors composed of nanowires as sensing material. Sensors 8:290–313

    Article  CAS  Google Scholar 

  6. Wei N, Chen JH, Zhang J, Wang K, Xu XW, Lin JH, Li GW, Lin XH, Chen YZ (2009) An electrochemical biosensor for detection of PML/RARA fusion gene using capture probe covalently immobilized onto poly-calcon carboxylic acid modified glassy carbon electrode. Talanta 78:1227–1234

    Article  CAS  Google Scholar 

  7. Lin XH, Wan HY, Zhang YF, Chen JH (2008) Studies of the interaction between Aloe-emodin and DNA and preparation of DNA biosensor for detection of PML-RAR alpha fusion gene in acute promyelocytic leukemia. Talanta 74:944–950

    Article  CAS  Google Scholar 

  8. Lai RY, Lagally ET, Lee SH, Soh HT, Plaxco KW, Heeger AJ (2006) Rapid, sequence-specific detection of unpurified PCR amplicons via a reusable, electrochemical sensor. Proc Natl Acad Sci USA 103:4017–4021

    Article  CAS  Google Scholar 

  9. Rebeca MC, de-los Patricia SA, Paulino TB (2007) Hairpin-DNA probe for enzyme-amplified electrochemical detection of Legionella pneumophila. Anal Chem 79:4050–4055

    Article  Google Scholar 

  10. Wang K, Sun ZL, Feng MJ, Liu AL, Yuan SY, Chen YZ, Lin XH (2011) Design of a sandwich-mode amperometric biosensor for detection of PML/RAR fusion gene using locked nucleic acids on gold electrode. Biosens Bioelectron 26:2870–2876

    Article  CAS  Google Scholar 

  11. Kierzek E, Pasternak A, Pasternak K, Gdaniec Z, Yildirim I, Turner D, Kierzek R (2009) Contributions of stacking, preorganization, and hydrogen bonding to the thermodynamic stability of duplexes between RNA and 2′-O-methyl RNA with locked nucleic acids. Biochemistry 48:4377–4387

    Article  CAS  Google Scholar 

  12. Stenvang J, Silahtaroglu AN, Lindow M, Elmen J, Kauppinen S (2008) The utility of LNA in microRNA-based cancer diagnostics and therapeutics. Semin Cancer Biol 18:89–102

    Article  CAS  Google Scholar 

  13. Steger G (1994) Thermal denaturation of double-stranded nucleic acids: prediction of temperatures critical for gradient gel electrophoresis and polymerase chain reaction. Nucl Acids Res 22:2760–2768

    Article  CAS  Google Scholar 

  14. Dubey RK, Tripathi DN (2005) Induction of resistance in host against the infection of leaf blight pathogen (Alternaria plaandui) in onion (Allium cepa var aggregatum). Indian J Biochem Biophys 42:301–307

    CAS  Google Scholar 

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Acknowledgments

The authors gratefully acknowledge the financial support of the National High Technology and Development of China (863 Project: 2012AA022604), the National Natural Science Foundation of China (21275028), the Fujian Provincial University-Industry Cooperation Science & Technology Major Program (2010Y4003), the Scientific Research Major Program of Fujian Medical University (09ZD013), the Natural Science Foundation of Fujian Province of China (2010 J05019), and the Foundation of Fujian Health Department (2010134).

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Authors and Affiliations

  1. Department of Pharmaceutical Analysis, Faculty of Pharmacy, Fujian Medical University, Fuzhou, 350004, China

    Yun Lei, Mei-juan Feng, Kun Wang, Li-qing Lin & Xin-hua Lin

  2. Fujian Institute of Hematology, The Affiliated Union Hospital of Fujian Medical University, Fuzhou, 350000, China

    Yuan-zhong Chen

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  1. Yun Lei
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  2. Mei-juan Feng
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  3. Kun Wang
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  4. Li-qing Lin
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  5. Yuan-zhong Chen
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  6. Xin-hua Lin
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Correspondence to Xin-hua Lin.

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Lei, Y., Feng, Mj., Wang, K. et al. Sequence-specific electrochemical detection of double-strand PCR amplicons of PML/RARα fusion gene in acute promyelocytic leukemia. Anal Bioanal Chem 405, 423–428 (2013). https://doi.org/10.1007/s00216-012-6477-6

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  • DOI: https://doi.org/10.1007/s00216-012-6477-6

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