**4.2. Invasive methods**

Methodologies to increase numbers of planktonic organisms that can be cultured from devices have been devised, using sonication of the devices [66, 67]. Metagenomic analysis of the results of next generation sequencing has been used to diagnose *A. defectiva* CNE [68]. A universal PCR/sequencing test has been applied to diagnose CNE on blood and valvular tissue [69]. Immunofluorescent antibody detection, Western blot analysis and real time-PCR of 16s RNA have been used to diagnose CNE due to *Bartonella* spp. [38]. PCR/ESI-MS has been utilized on valve tissue to diagnose CNE [70, 71].
