Polymerase chain reaction-based diagnosis of Mediterranean spotted fever in serum and tissue samples

• Published in: The American journal of tropical medicine and hygiene Vol. 67; no. 2; pp. 166 - 169
• Main Authors: Leitner, M, Yitzhaki, S, Rzotkiewicz, S, Keysary, A
• Format: Journal Article
• Language: English
• Published: Lawrence, KS ASTMH 01.08.2002; Allen Press
• Subjects: Bacterial diseases; Biological and medical sciences; Boutonneuse Fever - blood; Boutonneuse Fever - diagnosis; Boutonneuse Fever - microbiology; DNA Primers; DNA, Bacterial - analysis; Fluorescent Antibody Technique; Human bacterial diseases; Humans; Infectious diseases; Medical sciences; Polymerase Chain Reaction; Rickettsia conorii - genetics; Rickettsia conorii - isolation & purification; Rickettsia typhi - genetics; Rickettsia typhi - isolation & purification; Rickettsial diseases; Sensitivity and Specificity; Tropical bacterial diseases; Human; Rickettsialosis; Boutonneuse fever; Method; Rickettsia conorii; Infection; Rickettsiales; Polymerase chain reaction; Tissue; Rickettsial infection; Bacteriosis; Bacteria; Serum; Rickettsieae; Diagnosis; Molecular biology; Rickettsiaceae
• ISSN: 0002-9637; 1476-1645
• DOI: 10.4269/ajtmh.2002.67.166

A nested polymerase chain reaction (PCR) assay has been developed and used in the diagnosis of fatal and benign cases of Mediterranean spotted fever (MSF). The test was based on specific primers derived from a Rickettsia conorii 17-kD protein gene. A positive signal was obtained from spotted fever group (SFG) and typhus group (TG) rickettsiae. Discrimination between SFG and TG rickettsiae was based on a restriction fragment length polymorphism test. Other gram-negative bacterial species tested did not generate a signal, attesting for the specificity of the assay. The SFG-specific DNA fragment was detected in four of 29 acute-phase sera from serologically confirmed patients with MSF, while acute-phase sera from 25 patients without MSF were PCR negative. Acute-phase sera samples (five of five) and tissue autopsies (six of seven) from fatal suspected cases of MSF were PCR positive. The results demonstrate that sera and tissue samples are suitable specimens for the nested PCR tests, especially in fatal cases.