Entamoeba histolytica acetyl-CoA synthetase: biomarker of acute amoebic liver abscess.

Lim Boon Huat; Alfonso Olivos Garcia; Tan Zi Ning; Wong Weng Kin; Rahmah Noordin; Siti Shafiqah Anaqi Azham; Lee Zhi Jie; Guee Cher Ching; Foo Phiaw Chong; Pim Chau Dam

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Entamoeba histolytica acetyl-CoA synthetase: biomarker of acute amoebic liver abscess.

Author: Lim Boon HUAT ¹ ; Alfonso Olivos GARCIA ² ; Tan Zi NING ³ ; Wong Weng KIN ¹ ; Rahmah NOORDIN ⁴ ; Siti Shafiqah Anaqi AZHAM ¹ ; Lee Zhi JIE ¹ ; Guee Cher CHING ¹ ; Foo Phiaw CHONG ¹ ; Pim Chau DAM ¹
Author Information

1. Biomedicine Programme, School of Health Sciences, Universiti Sains Malaysia, 16150 Kubang Kerian, Kelantan, Malaysia.
2. Departmento de Medicina Experimental, Facultad de Medicina, Universidad Nacional Autonoma de Mexico, 04510 Mexico D.F., Mexico.
3. Department of Medicine, Allianze University College of Medical Sciences, Pulau Pinang, Malaysia.
4. Institute for Research in Molecular Medicine, 11800 Universiti Sains Malaysia, Penang, Malaysia.
Publication Type:Journal Article
Keywords: Acetyl-CoA synthetase; Amoebic liver abscess; Biomarker; Entamoeba histolytica; Recombinant EhACS; Western blot
From:Asian Pacific Journal of Tropical Biomedicine 2014;4(6):446-450
CountryChina
Language:English
Abstract:
OBJECTIVETo characterize the Entamoeba histolytica (E. histolytica) antigen(s) recognized by moribound amoebic liver abscess hamsters.
METHODSCrude soluble antigen of E. histolytica was probed with sera of moribund hamsters in 1D- and 2D-Western blot analyses. The antigenic protein was then sent for tandem mass spectrometry analysis. The corresponding gene was cloned and expressed in Escherichia coli BL21-AI to produce the recombinant E. histolytica ADP-forming acetyl-CoA synthetase (EhACS) protein. A customised ELISA was developed to evaluate the sensitivity and specificity of the recombinant protein.
RESULTSA ∼75 kDa protein band with a pI value of 5.91-6.5 was found to be antigenic; and not detected by sera of hamsters in the control group. Tandem mass spectrometry analysis revealed the protein to be the 77 kDa E. histolytica ADP-forming acetyl-CoA synthetase (EhACS). The customised ELISA results revealed 100% sensitivity and 100% specificity when tested against infected (n=31) and control group hamsters (n=5) serum samples, respectively.
CONCLUSIONSThis finding suggested the significant role of EhACS as a biomarker for moribund hamsters with acute amoebic liver abscess (ALA) infection. It is deemed pertinent that future studies explore the potential roles of EhACS in better understanding the pathogenesis of ALA; and in the development of vaccine and diagnostic tests to control ALA in human populations.