title: An alternative nested-PCR assay for the detection of Toxoplasma gondii strains based on GRA7 gene sequences.
abstract: Toxoplasma gondii is a widespread parasite able to infect virtually any nucleated cells of warm-blooded hosts. In some cases, T. gondii detection using already developed PCR primers can be inefficient in routine laboratory tests, especially to detect atypical strains. Here we report a new nested-PCR protocol able to detect virtually all T. gondii isolates. Analyzing 685 sequences available in GenBank, we determine that GRA7 is one of the most conserved genes of T. gondii genome. Based on an alignment of 85 GRA7 sequences new primer sets that anneal in the highly conserved regions of this gene were designed. The new GRA7 nested-PCR assay providing sensitivity and specificity equal to or greater than the gold standard PCR assays for T. gondii detection, that amplify the B1 sequence or the repetitive 529bp element.
authors.affiliation: Laboratório de Biologia Molecular Aplicada-LAPLIC, Departamento de Bioquímica, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Programa de Pós-Graduação em Biologia Parasitária, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Maria Eduarda S M
authors.initials: ME
authors.lastname: Costa
authors.affiliation: Laboratório de Biologia da Malária e Toxoplasmose, Departamento de Microbiologia e Parasitologia, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Claudio Bruno S
authors.initials: CB
authors.lastname: Oliveira
authors.affiliation: Laboratório de Biologia da Malária e Toxoplasmose, Departamento de Microbiologia e Parasitologia, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Joelma Maria de A
authors.initials: JM
authors.lastname: Andrade
authors.affiliation: Laboratório de Biologia da Malária e Toxoplasmose, Departamento de Microbiologia e Parasitologia, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Thatiany A
authors.initials: TA
authors.lastname: Medeiros
authors.affiliation: Laboratório de Biologia da Malária e Toxoplasmose, Departamento de Microbiologia e Parasitologia, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Programa de Pós-Graduação em Biologia Parasitária, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil.
authors.forename: Valter F Andrade
authors.initials: VF
authors.lastname: Neto
authors.affiliation: Laboratório de Biologia Molecular Aplicada-LAPLIC, Departamento de Bioquímica, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil; Programa de Pós-Graduação em Biologia Parasitária, Universidade Federal do Rio Grande do Norte, Natal, RN, Brazil. Electronic address: danielclanza@gmail.com.
authors.forename: Daniel C F
authors.initials: DC
authors.lastname: Lanza
mesh_terms: D000953:Antigens, Protozoan / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D015800:Protozoan Proteins / Q000235:genetics*; D012680:Sensitivity and Specificity; D014122:Toxoplasma / Q000235:genetics* / Q000302:isolation & purification*; D014123:Toxoplasmosis / Q000469:parasitology*
chemical_list: D000953:Antigens, Protozoan; D017931:DNA Primers; C112573:GRA7 protein, Toxoplasma gondii; D015800:Protozoan Proteins
keywords: 529bp region; Apicomplexa; B1 gene; Dense granule protein 7; Rhoptry; Toxoplasmosis