*5.3.3 Nested and semi-nested PCR*

In nested PCR, two sets of PCR primers are utilized to study the same genomic region. Currently, nested PCR and semi-nested PCR techniques are being developed to detect *Brucella* in human blood samples and are being tested in clinical settings for the diagnosis of human brucellosis. Lin et al. reported a nested PCR for the laboratory diagnosis of human brucellosis. During semi-nested PCR, two sets of PCR primers are used, but one of the primers in the second set is the same as that in the first [37].

## *5.3.4 Other PCR-based assays*

Polymerase chain reaction-enzyme immunoassay (PCR-EIA) was used by Vrioni et al. (2004) for the quick laboratory diagnosis of human brucellosis from peripheral blood. According to the findings, the PCR-EIA assay is a sensitive and specific technique that could help in the quick and precise diagnosis of acute human brucellosis [38].
