Evaluation of nested PCR for the diagnosis of scrub typhus among patients with acute pyrexia of unknown origin

A nested PCR technique was performed to detect a specific 483 bp DNA fragment of Orientia tsutsugamushi, the aetiological agent of scrub typhus, in 53 blood samples from 36 patients with acute pyrexia of unknown origin in southern Thailand. The specific primers could amplify the specific DNA from all 10 prototype strains of O. tsutsugamushi and all nine seropositive patients and three seronegative patients, while no DNA amplification was obtained with DNAs from other rickettsiae or from healthy persons or from patients with murine typhus. The specific PCR product was detectable in the blood for as long as 22 days after the onset of disease in patients without specific treatment and 27 days after receiving a single dose of doxycycline. Thus, nested PCR may be more sensitive than the serological test for diagnosis of scrub typhus and prolonged persistence of O. tsutsugamushi DNA in patients' blood was demonstrated despite clinical recovery of the patients.