RAPID-M™ technology: A cell-free method for protein microarray generation from PCR DNA

Suhanawati A. Nur, R. Suganti, K. Dzulaikha, M. A. Nor Azurah, H. Yuka, M. Nor Muhamad, Rahmah Mohd Amin, Sheila Nathan

Research output: Contribution to journalArticle

Abstract

Introduction: Burkholderia pseudomallei is the causative agent of melioidosis, an infectious disease with multifarious manifestations and has been listed as a potential biowarfare agent. The successful management of patients with severe melioidosis depends on early diagnosis and initiation of appropriate antibiotic therapy. Protein microarray has shown enormous potential for in vitro diagnostics due to miniaturization whilst generating a maximum of diagnostically relevant information from minute amount of sample. The Repeatable Arrays of Proteins using Immobilized DNA Microplates (RAPID-MTM) technology was developed to facilitate the development of high-throughput protein microarray platform that potentially contributes to the diagnostic approach. Objective: To develop protein microarray using RAPID-MTM technology for potential diagnostic applications. Methods: The Omp3 and GFP genes were immobilized onto beads during PCR amplification and these beads-immobilized DNA were then used as template for cell-free reactions. The synthesized proteins were printed directly onto glass slide using in situ protein purification to produce many identical copies of protein microarray. Result & Discussion: Expression of Omp3 and GFP proteins using beads-immobilized DNA yielded protein bands with expected molecular size of 27kDa and 30kDa, respectively as proven by the Western blot analysis. The Omp3 and GFP proteins were still functional after in situ purification on the Ni-NTA microarray slide. Conclusion: Protein microarray fabrication using RAPID-MTM technology has been successfully developed.

Original languageEnglish
Pages (from-to)242
Number of pages1
JournalAsian Pacific Journal of Tropical Disease
Volume4
Issue number3
DOIs
Publication statusPublished - Jun 2014

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Protein Array Analysis
Immobilized Nucleic Acids
Technology
Polymerase Chain Reaction
DNA
Melioidosis
Proteins
Biological Warfare Agents
Burkholderia pseudomallei
Miniaturization
Glass
Communicable Diseases
Early Diagnosis
Western Blotting
Anti-Bacterial Agents
Genes

ASJC Scopus subject areas

  • Infectious Diseases
  • Microbiology (medical)

Cite this

RAPID-M™ technology : A cell-free method for protein microarray generation from PCR DNA. / Nur, Suhanawati A.; Suganti, R.; Dzulaikha, K.; Nor Azurah, M. A.; Yuka, H.; Nor Muhamad, M.; Mohd Amin, Rahmah; Nathan, Sheila.

In: Asian Pacific Journal of Tropical Disease, Vol. 4, No. 3, 06.2014, p. 242.

Research output: Contribution to journalArticle

Nur, SA, Suganti, R, Dzulaikha, K, Nor Azurah, MA, Yuka, H, Nor Muhamad, M, Mohd Amin, R & Nathan, S 2014, 'RAPID-M™ technology: A cell-free method for protein microarray generation from PCR DNA', Asian Pacific Journal of Tropical Disease, vol. 4, no. 3, pp. 242. https://doi.org/10.1016/S2222-1808(14)60548-X
Nur, Suhanawati A. ; Suganti, R. ; Dzulaikha, K. ; Nor Azurah, M. A. ; Yuka, H. ; Nor Muhamad, M. ; Mohd Amin, Rahmah ; Nathan, Sheila. / RAPID-M™ technology : A cell-free method for protein microarray generation from PCR DNA. In: Asian Pacific Journal of Tropical Disease. 2014 ; Vol. 4, No. 3. pp. 242.
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