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

BMC Infectious Diseases

Qasem Asgari¹  Farah Bokharaei-Salim²  Sayyed Amirpooya Alemzadeh³  Alireza Hedayatfar³  Saba Garshasbi⁴  Khadijeh Khanaliha⁵  Abdoulreza Esteghamati⁵  Borna Salemi⁶ 

[1] Department of Parasitology and Mycology, School of Medicine, Shiraz University of Medical Sciences;Department of Virology, School of Medicine, Iran University of Medical Sciences;Eye Research Center, The Five Senses Institute, Iran University of Medical Sciences;Iran University of Medical Sciences;Research Center of Pediatric Infectious Diseases, Institute of Immunology and Infectious Diseases, Iran University of Medical Sciences;Student Research Committee, School of Medicine, Iran University of Medical Sciences;

关键词: Toxoplasma gondii;    Retinochoroiditis;    Real-time PCR;    Ocular toxoplasmosis;    PCR;    Peripheral blood mononuclear cells;   

DOI  :  10.1186/s12879-021-06873-3

来源: DOAJ

【 摘 要 】

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.

【 授权许可】

Unknown