Comparison of the sensitivity and cross-reactivity between heat-killed and formalin-killed cells during the development of indirect competitive ELISA for the detection of Salmonella Typhimurium

Authors

  • Noor Azlina Masdor Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • Y. Noor Aseha Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • K. Noorzila Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • Mohd Yunus Abd Shukor Department of Biochemistry, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, UPM 43400 Serdang, Selangor, Malaysia.
  • R.I. Mohammad Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia
  • I. Zamri Biotechnology Research Centre, MARDI, P. O. Box 12301, 50774 Kuala Lumpur, Malaysia

DOI:

https://doi.org/10.54987/jebat.v2i1.144

Keywords:

indirect ELISA, Salmonella Typhimurium, formalin-killed, heat-killed, four parameter logistic equation

Abstract

Newer discovery of Salmonella strains warranted the development of newer ELISA system. The
aim of this works is to develop a sensitive and simple indirect ELISA for the detection of
Salmonella Typhimurium using formalin- and heat-killed cells as immunogens. The parameters of
the resultant calibration curve for the formalin-killed cells (FKC) were minimum absorbance of
0.101± 0.011, maximum absorbance of 2.804± 0.020, slope parameter of 1.197, while c, the
concentration that results in 50% response was 4.22 x 106 CFU/ml. The LOD and LOQ were
19,300 and 52,700 CFU/mL, respectively. The parameters of the typical calibration curve for
heat-killed cells (HKC) were minimum absorbance of 0.115± 0.013, maximum absorbance of
2.810± 0.021, slope parameter was 1.441, while c, the concentration that results in 50% response
was 2.15 x 106 CFU/ml. The LOD and LOQ were 47,100 and 123,000 CFU/mL, respectively.
Antibody raised to FKC was twice the sensitivity and approximately ten times the linear range
compared to antibody raised to HKC. Antibody produced from formalin-killed cells exhibited
lower cross reactivity than heat-killed cells. The results of this work could open new avenues on
optimization of antigen preparation for maximal sensitivity of the developed polyclonal antibody.

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Published

2014-07-31

How to Cite

Masdor, N. A., Noor Aseha, Y., Noorzila, K., Shukor, M. Y. A., Mohammad, R., & Zamri, I. (2014). Comparison of the sensitivity and cross-reactivity between heat-killed and formalin-killed cells during the development of indirect competitive ELISA for the detection of Salmonella Typhimurium. Journal of Environmental Bioremediation and Toxicology, 2(1), 23–27. https://doi.org/10.54987/jebat.v2i1.144

Issue

Vol. 2 No. 1 (2014)

Section

Articles

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