Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients

Abstract Backgrounds PCR is a proper technique that significantly improves toxoplasmosis diagnosis. However, a more sensitive technique is required. This study compared real-time PCR with nested PCR using B1, SAG-4, and MAG-1 bradyzoite genes to diagnose toxoplasmosis in toxoplasmic retinochoroiditi...

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Autores principales:	Khadijeh Khanaliha, Farah Bokharaei-Salim, Alireza Hedayatfar, Abdoulreza Esteghamati, Sayyed Amirpooya Alemzadeh, Qasem Asgari, Saba Garshasbi, Borna Salemi
Formato:	article
Lenguaje:	EN
Publicado:	BMC 2021
Materias:	Toxoplasma gondii Retinochoroiditis Real-time PCR Ocular toxoplasmosis PCR Peripheral blood mononuclear cells Infectious and parasitic diseases RC109-216
Acceso en línea:	https://doaj.org/article/b8d5bd04767243f4833cffd568dfb2a5
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spelling	oai:doaj.org-article:b8d5bd04767243f4833cffd568dfb2a52021-11-28T12:41:45ZComparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients10.1186/s12879-021-06873-31471-2334https://doaj.org/article/b8d5bd04767243f4833cffd568dfb2a52021-11-01T00:00:00Zhttps://doi.org/10.1186/s12879-021-06873-3https://doaj.org/toc/1471-2334Abstract Backgrounds PCR is a proper technique that significantly improves toxoplasmosis diagnosis. However, a more sensitive technique is required. This study compared real-time PCR with nested PCR using B1, SAG-4, and MAG-1 bradyzoite genes to diagnose toxoplasmosis in toxoplasmic retinochoroiditis patients. Methods Blood samples were collected from 10 patients with active toxoplasmic chorioretinal lesions and 10 healthy individuals. Blood samples including peripheral blood mononuclear cells (PBMCs), serum and whole blood samples were used for DNA extraction. Serum was also used to detect anti-toxoplasma IgG and IgM antibodies. Nested PCR and real-time PCR were performed using B1, SAG-4, and MAG-1 target genes. Results Five (50%) out of the 10 patients were tested positive for toxoplasmosis with nested PCR using the PBMC samples. All the five patients tested positive with nested PCR were also tested positive for toxoplasmosis with real-time PCR using the PBMC samples. The real-time PCR results demonstrated that 9(90%) out of the 10 patients were positive based on B1 and the remaining one (10%) was positive only based on MAG-1. In general, of the patients, five (50%) were positive using SAG-4 and three (30%) were positive in term of MAG-1 using PBMCs with real-time PCR. Conclusion It appears that PBMC samples have the best performance as the PCR extraction method and are a good source for toxoplasmosis diagnosis. The use of B22 and B23 target genes due to their high sensitivity and specificity along with bradyzoite genes are recommended for toxoplasmosis diagnosis using PBMC samples with real-time PCR.Khadijeh KhanalihaFarah Bokharaei-SalimAlireza HedayatfarAbdoulreza EsteghamatiSayyed Amirpooya AlemzadehQasem AsgariSaba GarshasbiBorna SalemiBMCarticleToxoplasma gondiiRetinochoroiditisReal-time PCROcular toxoplasmosisPCRPeripheral blood mononuclear cellsInfectious and parasitic diseasesRC109-216ENBMC Infectious Diseases, Vol 21, Iss 1, Pp 1-7 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Toxoplasma gondii Retinochoroiditis Real-time PCR Ocular toxoplasmosis PCR Peripheral blood mononuclear cells Infectious and parasitic diseases RC109-216
spellingShingle	Toxoplasma gondii Retinochoroiditis Real-time PCR Ocular toxoplasmosis PCR Peripheral blood mononuclear cells Infectious and parasitic diseases RC109-216 Khadijeh Khanaliha Farah Bokharaei-Salim Alireza Hedayatfar Abdoulreza Esteghamati Sayyed Amirpooya Alemzadeh Qasem Asgari Saba Garshasbi Borna Salemi Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
description	Abstract Backgrounds PCR is a proper technique that significantly improves toxoplasmosis diagnosis. However, a more sensitive technique is required. This study compared real-time PCR with nested PCR using B1, SAG-4, and MAG-1 bradyzoite genes to diagnose toxoplasmosis in toxoplasmic retinochoroiditis patients. Methods Blood samples were collected from 10 patients with active toxoplasmic chorioretinal lesions and 10 healthy individuals. Blood samples including peripheral blood mononuclear cells (PBMCs), serum and whole blood samples were used for DNA extraction. Serum was also used to detect anti-toxoplasma IgG and IgM antibodies. Nested PCR and real-time PCR were performed using B1, SAG-4, and MAG-1 target genes. Results Five (50%) out of the 10 patients were tested positive for toxoplasmosis with nested PCR using the PBMC samples. All the five patients tested positive with nested PCR were also tested positive for toxoplasmosis with real-time PCR using the PBMC samples. The real-time PCR results demonstrated that 9(90%) out of the 10 patients were positive based on B1 and the remaining one (10%) was positive only based on MAG-1. In general, of the patients, five (50%) were positive using SAG-4 and three (30%) were positive in term of MAG-1 using PBMCs with real-time PCR. Conclusion It appears that PBMC samples have the best performance as the PCR extraction method and are a good source for toxoplasmosis diagnosis. The use of B22 and B23 target genes due to their high sensitivity and specificity along with bradyzoite genes are recommended for toxoplasmosis diagnosis using PBMC samples with real-time PCR.
format	article
author	Khadijeh Khanaliha Farah Bokharaei-Salim Alireza Hedayatfar Abdoulreza Esteghamati Sayyed Amirpooya Alemzadeh Qasem Asgari Saba Garshasbi Borna Salemi
author_facet	Khadijeh Khanaliha Farah Bokharaei-Salim Alireza Hedayatfar Abdoulreza Esteghamati Sayyed Amirpooya Alemzadeh Qasem Asgari Saba Garshasbi Borna Salemi
author_sort	Khadijeh Khanaliha
title	Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
title_short	Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
title_full	Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
title_fullStr	Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
title_full_unstemmed	Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
title_sort	comparison of real-time pcr and nested pcr for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients
publisher	BMC
publishDate	2021
url	https://doaj.org/article/b8d5bd04767243f4833cffd568dfb2a5
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Comparison of real-time PCR and nested PCR for toxoplasmosis diagnosis in toxoplasmic retinochoroiditis patients

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