| Autor: |
Pacce, Violetta Dias, Souza, Margarida Neves, de Oliveira, Natasha Rodrigues, Kremer, Frederico Schmitt, Jorge, Sérgio, Ikuta, Nilo, Lunge, Vagner Ricardo, Dellagostin, Odir Antônio |
| Zdroj: |
Brazilian Journal of Microbiology; June 2022, Vol. 53 Issue: 2 p1029-1037, 9p |
| Abstrakt: |
Leptospirosis is a zoonotic disease caused by pathogenic species of Leptospira. Due to the similarity with clinical signs of other febrile diseases, early diagnosis remains challenging. Real-time PCR has been used for direct detection of Leptospira, but it requires thermocyclers and highly trained personnel. Loop-mediated isothermal amplification (LAMP) is a simple and rapid DNA-based assay. Therefore, here we have developed PCR and LAMP assays targeting two novel genes, lic13162and lic20239, and also lipL32gene to detect pathogenic Leptospira. Analytical and diagnostic performances were compared with bacterial isolates (including different Leptospiraspecies and serovars) and clinical samples. The results demonstrated that PCR assays targeting lic13162and lic20239were successful to amplify Leptospira, but LAMP not. However, both PCR and LAMP targeting lipL32could detect pathogenic Leptospira. LAMP lipL32could be performed in 30 min with a detection limit of 156 cells/mL. Diagnostic performance of lipL32-LAMP presented 84.2% sensitivity and 93.2% specificity. In conclusion, lipL32PCR and LAMP are effective methods to detect pathogenic Leptospiradirectly from clinical samples. |
| Databáze: |
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