TY - JOUR
T1 - Application of recombinant SMR-domain containing protein of angiostrongylus cantonensis in immunoblot diagnosis of human angiostrongyliasis
AU - Vitta, Apichat
AU - Yoshino, Timothy P.
AU - Kalambaheti, Thareerat
AU - Komalamisra, Chalit
AU - Waikagul, Jitra
AU - Ruangsittichai, Jiraporn
AU - Dekumyoy, Paron
PY - 2010/7
Y1 - 2010/7
N2 - The aim of this study was to find novel proteins expressed from an Angiostrongylus cantonensis adult female worm cDNA library for serodiagnosis of angiostrongyliasis. An immuno-dominant clone, fAC22, was identified by immunoscreening with pooled positive sera from proven angiostrongyliasis patients. The clone contained an open reading frame of 2,136 bp encoding a 80.5 kDa protein with a predicted isoelectric point of 5.8. The deduced amino acid sequence (712 amino acids) contained the conserved domain of Small mutS related (Smr) superfamily protein, with similarity with the Smr domain protein of Brugia malayi. The fusion His-tagged 81 kDa recombinant protein expressed as inclusion body in Escherichia coli was solubilized and purified by Ni-affinity chromatography for use in immunoblot analysis. Its sensitivity, specificity, positive and negative predictive values in immunodiagnostic test was 93.5, 91.5, 79.0 and 97.5%, respectively. Although some cross-reactivity of the antigen was observed among gnathostomiasis, bancroftian filariasis, ascariasis, echinococcosis, paragonimiasis and opisthorchiasis, sera from 14 other infections were all negative. These data indicate its possible application in immunodiagnosis of clinically suspected angiostrongyliasis.
AB - The aim of this study was to find novel proteins expressed from an Angiostrongylus cantonensis adult female worm cDNA library for serodiagnosis of angiostrongyliasis. An immuno-dominant clone, fAC22, was identified by immunoscreening with pooled positive sera from proven angiostrongyliasis patients. The clone contained an open reading frame of 2,136 bp encoding a 80.5 kDa protein with a predicted isoelectric point of 5.8. The deduced amino acid sequence (712 amino acids) contained the conserved domain of Small mutS related (Smr) superfamily protein, with similarity with the Smr domain protein of Brugia malayi. The fusion His-tagged 81 kDa recombinant protein expressed as inclusion body in Escherichia coli was solubilized and purified by Ni-affinity chromatography for use in immunoblot analysis. Its sensitivity, specificity, positive and negative predictive values in immunodiagnostic test was 93.5, 91.5, 79.0 and 97.5%, respectively. Although some cross-reactivity of the antigen was observed among gnathostomiasis, bancroftian filariasis, ascariasis, echinococcosis, paragonimiasis and opisthorchiasis, sera from 14 other infections were all negative. These data indicate its possible application in immunodiagnosis of clinically suspected angiostrongyliasis.
KW - Angiostrongylus cantonensis
KW - Eosinophilic meningitis
KW - Immunodiagnosis
KW - Recombinant fusion protein
UR - http://www.scopus.com/inward/record.url?scp=79952113575&partnerID=8YFLogxK
M3 - Article
C2 - 21073053
AN - SCOPUS:79952113575
SN - 0125-1562
VL - 41
SP - 785
EP - 799
JO - Southeast Asian Journal of Tropical Medicine and Public Health
JF - Southeast Asian Journal of Tropical Medicine and Public Health
IS - 4
ER -