Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera

Atefeh Amerizadeh, Zulkarnain Md Idris, Boon Yin Khoo, Dupadahalli Kotresha, Muhammad Hafiznur Yunus, Izzati Zahidah Abdul Karim, Geita Saadatnia, Ai Ying Teh, Rahmah Noordin

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Toxoplasmosis is an infection caused by the parasite Toxoplasma gondii. Chronically-infected individuals with a compromised immune system are at risk for reactivation of the disease. In-vivo induced antigen technology (IVIAT) is a promising method for the identification of antigens expressed in-vivo. The aim of the present study was to apply IVIAT to identify antigens which are expressed in-vivo during T. gondii infection using sera from individuals with chronic toxoplasmosis. Forty serum samples were pooled, pre-adsorped against three different preparations of antigens, from each in-vitro grown T. gondii and Escherichia coli XLBlue MRF', and then used to screen a T. gondii cDNA expression library. Sequencing of DNA inserts from positive clones showed eight open reading frames with high homology to T. gondii genes. Expression analysis using quantitative real-time PCR showed that SAG1-related sequence 3 (SRS3) and two hypothetical genes were up-regulated in-vivo relative to their expression levels in-vitro. These three proteins also showed high sensitivity and specificity when tested with individual serum samples. Five other proteins namely M16 domain peptidase, microneme protein, elongation factor 1-alpha, pre-mRNA-splicing factor and small nuclear ribonucleoprotein F had lower RNA expression in-vivo as compared to in-vitro. SRS3 and the two hypothetical proteins warrant further investigation into their roles in the pathogenesis of toxoplasmosis.

Original languageEnglish
Pages (from-to)60-66
Number of pages7
JournalMicrobial Pathogenesis
Volume54
Issue number1
DOIs
Publication statusPublished - Jan 2013
Externally publishedYes

Fingerprint

Toxoplasmosis
Toxoplasma
Gene Library
Antigens
Serum
Proteins
Small Nuclear Ribonucleoproteins
Peptide Elongation Factor 1
Technology
Parasitic Diseases
RNA Precursors
DNA Sequence Analysis
Open Reading Frames
Genes
Real-Time Polymerase Chain Reaction
Immune System
Peptide Hydrolases
Clone Cells
RNA
Escherichia coli

Keywords

  • Chronic sera
  • In-vivo induced antigens
  • Real-time PCR
  • Toxoplasma gondii

ASJC Scopus subject areas

  • Microbiology
  • Infectious Diseases

Cite this

Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera. / Amerizadeh, Atefeh; Md Idris, Zulkarnain; Khoo, Boon Yin; Kotresha, Dupadahalli; Yunus, Muhammad Hafiznur; Abdul Karim, Izzati Zahidah; Saadatnia, Geita; Teh, Ai Ying; Noordin, Rahmah.

In: Microbial Pathogenesis, Vol. 54, No. 1, 01.2013, p. 60-66.

Research output: Contribution to journalArticle

Amerizadeh, A, Md Idris, Z, Khoo, BY, Kotresha, D, Yunus, MH, Abdul Karim, IZ, Saadatnia, G, Teh, AY & Noordin, R 2013, 'Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera', Microbial Pathogenesis, vol. 54, no. 1, pp. 60-66. https://doi.org/10.1016/j.micpath.2012.09.006
Amerizadeh, Atefeh ; Md Idris, Zulkarnain ; Khoo, Boon Yin ; Kotresha, Dupadahalli ; Yunus, Muhammad Hafiznur ; Abdul Karim, Izzati Zahidah ; Saadatnia, Geita ; Teh, Ai Ying ; Noordin, Rahmah. / Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera. In: Microbial Pathogenesis. 2013 ; Vol. 54, No. 1. pp. 60-66.
@article{da893d40c333488f9023aa104b407ede,
title = "Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera",
abstract = "Toxoplasmosis is an infection caused by the parasite Toxoplasma gondii. Chronically-infected individuals with a compromised immune system are at risk for reactivation of the disease. In-vivo induced antigen technology (IVIAT) is a promising method for the identification of antigens expressed in-vivo. The aim of the present study was to apply IVIAT to identify antigens which are expressed in-vivo during T. gondii infection using sera from individuals with chronic toxoplasmosis. Forty serum samples were pooled, pre-adsorped against three different preparations of antigens, from each in-vitro grown T. gondii and Escherichia coli XLBlue MRF', and then used to screen a T. gondii cDNA expression library. Sequencing of DNA inserts from positive clones showed eight open reading frames with high homology to T. gondii genes. Expression analysis using quantitative real-time PCR showed that SAG1-related sequence 3 (SRS3) and two hypothetical genes were up-regulated in-vivo relative to their expression levels in-vitro. These three proteins also showed high sensitivity and specificity when tested with individual serum samples. Five other proteins namely M16 domain peptidase, microneme protein, elongation factor 1-alpha, pre-mRNA-splicing factor and small nuclear ribonucleoprotein F had lower RNA expression in-vivo as compared to in-vitro. SRS3 and the two hypothetical proteins warrant further investigation into their roles in the pathogenesis of toxoplasmosis.",
keywords = "Chronic sera, In-vivo induced antigens, Real-time PCR, Toxoplasma gondii",
author = "Atefeh Amerizadeh and {Md Idris}, Zulkarnain and Khoo, {Boon Yin} and Dupadahalli Kotresha and Yunus, {Muhammad Hafiznur} and {Abdul Karim}, {Izzati Zahidah} and Geita Saadatnia and Teh, {Ai Ying} and Rahmah Noordin",
year = "2013",
month = "1",
doi = "10.1016/j.micpath.2012.09.006",
language = "English",
volume = "54",
pages = "60--66",
journal = "Microbial Pathogenesis",
issn = "0882-4010",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Identification of Toxoplasma gondii in-vivo induced antigens by cDNA library immunoscreening with chronic toxoplasmosis sera

AU - Amerizadeh, Atefeh

AU - Md Idris, Zulkarnain

AU - Khoo, Boon Yin

AU - Kotresha, Dupadahalli

AU - Yunus, Muhammad Hafiznur

AU - Abdul Karim, Izzati Zahidah

AU - Saadatnia, Geita

AU - Teh, Ai Ying

AU - Noordin, Rahmah

PY - 2013/1

Y1 - 2013/1

N2 - Toxoplasmosis is an infection caused by the parasite Toxoplasma gondii. Chronically-infected individuals with a compromised immune system are at risk for reactivation of the disease. In-vivo induced antigen technology (IVIAT) is a promising method for the identification of antigens expressed in-vivo. The aim of the present study was to apply IVIAT to identify antigens which are expressed in-vivo during T. gondii infection using sera from individuals with chronic toxoplasmosis. Forty serum samples were pooled, pre-adsorped against three different preparations of antigens, from each in-vitro grown T. gondii and Escherichia coli XLBlue MRF', and then used to screen a T. gondii cDNA expression library. Sequencing of DNA inserts from positive clones showed eight open reading frames with high homology to T. gondii genes. Expression analysis using quantitative real-time PCR showed that SAG1-related sequence 3 (SRS3) and two hypothetical genes were up-regulated in-vivo relative to their expression levels in-vitro. These three proteins also showed high sensitivity and specificity when tested with individual serum samples. Five other proteins namely M16 domain peptidase, microneme protein, elongation factor 1-alpha, pre-mRNA-splicing factor and small nuclear ribonucleoprotein F had lower RNA expression in-vivo as compared to in-vitro. SRS3 and the two hypothetical proteins warrant further investigation into their roles in the pathogenesis of toxoplasmosis.

AB - Toxoplasmosis is an infection caused by the parasite Toxoplasma gondii. Chronically-infected individuals with a compromised immune system are at risk for reactivation of the disease. In-vivo induced antigen technology (IVIAT) is a promising method for the identification of antigens expressed in-vivo. The aim of the present study was to apply IVIAT to identify antigens which are expressed in-vivo during T. gondii infection using sera from individuals with chronic toxoplasmosis. Forty serum samples were pooled, pre-adsorped against three different preparations of antigens, from each in-vitro grown T. gondii and Escherichia coli XLBlue MRF', and then used to screen a T. gondii cDNA expression library. Sequencing of DNA inserts from positive clones showed eight open reading frames with high homology to T. gondii genes. Expression analysis using quantitative real-time PCR showed that SAG1-related sequence 3 (SRS3) and two hypothetical genes were up-regulated in-vivo relative to their expression levels in-vitro. These three proteins also showed high sensitivity and specificity when tested with individual serum samples. Five other proteins namely M16 domain peptidase, microneme protein, elongation factor 1-alpha, pre-mRNA-splicing factor and small nuclear ribonucleoprotein F had lower RNA expression in-vivo as compared to in-vitro. SRS3 and the two hypothetical proteins warrant further investigation into their roles in the pathogenesis of toxoplasmosis.

KW - Chronic sera

KW - In-vivo induced antigens

KW - Real-time PCR

KW - Toxoplasma gondii

UR - http://www.scopus.com/inward/record.url?scp=84871175112&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84871175112&partnerID=8YFLogxK

U2 - 10.1016/j.micpath.2012.09.006

DO - 10.1016/j.micpath.2012.09.006

M3 - Article

C2 - 23044055

AN - SCOPUS:84871175112

VL - 54

SP - 60

EP - 66

JO - Microbial Pathogenesis

JF - Microbial Pathogenesis

SN - 0882-4010

IS - 1

ER -