ELISA-based colorimetric detection of Rickettsia tsutsugamushi DNA from patient sera by nested polymerase chain reaction.

ABSTRACT

A rapid diagnostic system for scrub typhus was established using colorimetric detection of nested polymerase chain reaction (PCR). This system relied on binding the amplified DNA via a sequence in one of oligodeoxyribonucleotide to the DNA-binding protein GCN4 coated on the well of a micotiter dish. The primer pairs used for the nested PCR were designed on the basis of the homologous nucleotide sequence of the gene that encodes the 56 kDa antigen of serovariants. With this colorimetric PCR, diagnosis can be performed easily from serum samples of patients before the antibody titer increases or in the early stage of the disease. Furthermore, these positive results are able to be confirmed by pathogenic isolation.