Abstract
Although the excretory-secretory (ES) proteins of Trichinella spiralis muscle larvae are the most commonly used diagnostic antigens for trichinellosis, their main disadvantage is the false negative results during the early stage of infection and cross-reaction of their main components (43, 45, 49, and 53 kDa) with sera of patients with other helminthiasis. The aim of this study was to identify early specific diagnostic antigens in T. spiralis ES proteins with 30–40 kDa. The ES proteins were analyzed by two-dimensional electrophoresis (2-DE), and a total of approximately 150 proteins spots were detected with isoelectric point (pI) varying from 4 to 7 and molecular weight from 14 to 66 kDa. When probed with sera from infected mice at 18 days postinfection, ten protein spots with molecular weight of 30–40 kDa were recognized and identified by MALDI-TOF/TOF-MS. All of ten spots were successfully identified and characterized to correlate with five different proteins, including two potential serine proteases, one antigen targeted by protective antibodies, one deoxyribonuclease (DNase) II, and one conserved hypothetical protein. These proteins might be the early specific diagnostic antigens for trichinellosis.
Highlights
Trichinellosis remains an important food-borne parasitic zoonosis in many parts of the world [1, 2]
The ES antigens of T. spiralis muscle larvae (ML) were recommended to be used in Western blot for detecting anti-Trichinella antibodies by the International Commission on Trichinellosis (ICT) [10], and their main components were 43, 45, 49, and 53 kDa proteins analyzed by SDS-PAGE or two-dimensional electrophoresis (2-DE) gel analysis [11, 12]
The risk of cross-reactions using ES proteins is low in industrialized countries, cross-reactions do occur with anisakiasis or other larval migrans of unknown species
Summary
Trichinellosis remains an important food-borne parasitic zoonosis in many parts of the world [1, 2]. The ES antigens of T. spiralis ML were recommended to be used in Western blot for detecting anti-Trichinella antibodies by the International Commission on Trichinellosis (ICT) [10], and their main components were 43, 45, 49, and 53 kDa proteins analyzed by SDS-PAGE or two-dimensional electrophoresis (2-DE) gel analysis [11, 12] These main components of T. spiralis ES proteins were usually cross-reacted with sera of patients with echinococcosis, cysticercosis, schistosomiasis, paragonimiasis, and clonorchiasis [9, 13,14,15]. This is important in developing countries where human helminth infections are common and cross-reactions with these parasites could give false positive results [16]. The main immunoreactive protein spots with 30–40 kDa were identified and characterized by Matrix-assisted-laser-desorptionionization- (MALDI-) time-of-flight (TOF)/TOF-MS analyses in combination with bioinformatics analysis
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