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Vol. 113, Issue 2, February 2010, pp. 115-126

 

Bullet

 

Development of Piezoelectric DNA-Based Biosensor for Direct Detection of Mycobacterium Tuberculosis in Clinical Specimens

 

1Thongchai KAEWPHINIT, 2Somchai SANTIWATANAKUL, 3Chamras PROMPTMAS and 1Kosum CHANSIRI

1Department of Biochemistry, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand

Tel.: +66 2260-2122 ext 4605, fax: +66 2260-0125.

E-mails: tkaewphinit@yahoo.com, kosum@swu.ac.th

2Department of Pathology, Faculty of Medicine, Srinakharinwirot University, Sukhumvit 23, Bangkok 10110, Thailand

E-mail: titi41@yahoo.com

3Department of Clinical Chemistry, Faculty of Medical Technology, Mahidol University, Prannok, Bangkok, 10700, Thailand

E-mail: mtcpm@mahidol.ac.th

 

 

Received: 26 December 2009   /Accepted: 19 February 2010   /Published: 26 February 2010

 

Abstract: This study was focused on establishment of piezoelectric biosensor for direct detection of Mycobacterium tuberculosis (MTB) in clinical specimens. The quartz crystal immobilized via 3-mercaptopropionic acid (MPA)/avidin/DNA biotinylated probe on gold surface and hybridization of the DNA target to DNA biotinylated probe. The optimal concentration of MPA, avidin and 5-biotinylated DNA probe for immobilization of specific DNA probe on gold surface were 15 mM, 0.1 mg/ml and 1.5 μM, respectively. The detection of genomic DNA digestion in the range from 0.5 to 30 μg/ml. The fabricated biosensor was evaluated through an examination of 200 samples. No cross hybridization were observed against M. avium complex (MAC) and other microorganism. This target DNA preparation without amplification will reduce time consuming, costs, and the tedious step of amplification. This study can be extended to develop the new method which is high sensitivity, specificity, cheap, easy to use, and rapid for detection of MTB in many fields.

 

Keywords: Mycobacterium tuberculosis, Piezoelectric biosensor, Hybridization, Digestion genomic DNA

 

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