BactoReal® Kit Leptospira spp. Multiplex (16S+LipL32) is a multiplex real-time PCR assay for detection and differentiation of DNA of pathogenic and intermediately pathogenic Leptospira species. This test was developed and validated for the ABI PRISM® 7500 (Fast) instrument (Life Technologies), LightCycler® 480 (Roche) and Mx3005P® (Agilent), but is also suitable for other real-time PCR instruments. The test facilitates the rapid and sensitive detection of DNA of pathogenic and intermediately pathogenic Leptospira species from samples purified from blood, urine or kidney tissue. Leptospira DNA can be recovered efficiently from sample material using the QIAamp DNA Mini Kit extraction methods, for example.