A quantitative assay method of Toxoplasma gondii HSP70 mRNA by quantitative competitive-reverse transcriptase-PCR

• Published in: Parasitology international 2004-03, Vol.53 (1), p.49-58
• Main Authors: Piao, Lian-Xun, Aosai, Fumie, Chen, Mei, Fang, Hao, Mun, Hye-Seong, Norose, Kazumi, Yano, Akihiko
• Format: Article
• Language: English
• Subjects: Animals; Danger signal; HSP70 Heat-Shock Proteins - genetics; HSP70 Heat-Shock Proteins - metabolism; Interferon-gamma - genetics; Liver Function Tests; Mice; Mice, Inbred BALB C; Mice, Knockout; Protozoan Proteins - genetics; Protozoan Proteins - metabolism; QC-RT-PCR; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - genetics; RNA, Messenger - metabolism; T.g.HSP70; tachyzoites; Toxoplasma - genetics; Toxoplasma - growth & development; Toxoplasma - metabolism; Toxoplasma - pathogenicity; Toxoplasma gondii; Toxoplasmosis, Animal - mortality; Toxoplasmosis, Animal - parasitology; Virulence
• ISSN: 1383-5769; 1873-0329
• DOI: 10.1016/j.parint.2003.11.001

Toxoplasma gondii ( T. gondii)-derived heat shock protein 70 ( T.g.HSP70) has been identified as a virulent molecule expressing only in T. gondii tachyzoites during lethal acute infection. Therefore, it is of importance to determine the expression of T.g.HSP70 mRNA in a quantitative manner for analysis of virulence of T. gondii in tissues. We have constructed a competitor T.g.HSP70 and have successfully established a quantitative competitive-reverse transcriptase–polymerase chain reaction (QC-RT-PCR) targeting T.g.HSP70 gene. By using the established QC-RT-PCR method, we have demonstrated that the copy number of T.g.HSP70 mRNA per T. gondii tachyzoite was highest in the lung among the organs examined in interferon-γ knockout (GKO) mice.