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Published on: November 30, 2022
[Spotted fever group rickettsiosis and vectors in Kanagawa prefecture]
T Katayama1, Y Furuya, Y Yoshida
1Kanagawa Prefectural Public Health Laboratory, Japan.
Abstract:
Primer pairs for PCR were designed from the gene encoding the 17,000-molecular-weight genus-common antigen of Rickettsia japonica, Rickettsia rickettsii, Rickettsia conorii, Rickettsia typhi and Rickettsia prowazekii. Primers R1, R2 were designed for amplifying the genomic DNA from spotted fever group (SFG) rickettsiae and epidemic typhus rickettsiae. Primers Rj5, Rj10 were designed for amplifying the genomic DNA from only R. japonica. Using the primers R1, R2, about a 540-bp fragment was observed by amplifying the genomic DNA from R. japonica, R. rickettsii, R. conorii, Thai tick typhus TT-118, Rickettsia sibirica, Rickettsia montana, Rickettsia askari, R. typhi, R. prowazekii and Katayama strain isolated from the patient infected with SFG rickettsiae. Using the primers Rj5, Rj10, the 357-bp fragment was observed by amplifying the genomic DNA from R. japonica and Katayama strain. Therefore, the Katayama strain was identified to belong to R. japonica. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp bands were amplified from blood of the patients infected with SFG rickettsiae in Kanagawa prefecture. These findings indicate that the causative agent of SFG rickettsiosis in these two patients was R. japonica. The ticks, Ixodes ovatus and Haemaphysalis flava, were collected by out field research in Kanagawa prefecture. With primers R1, R2 and Rj5, Rj10, 537 bp and 357 bp were amplified from these ticks. This indicates that I. ovatus and H. flava were the vector of R. japonica in Kanagawa prefecture. Also, with the primers R1, R2, about a 540 bp fragment was amplified but with primers Rj5, Rj10, no fragments were amplified from I. ovatus and H. flava. Therefore, these ticks may have SFG rickettsiae other than R. japonica and epidemic typhus rickettsiae.
Insights
New PCR primers accurately identify Rickettsia japonica, the cause of spotted fever group rickettsiosis. These primers also identified R. japonica in ticks, confirming them as vectors in Kanagawa prefecture.
Area of Science:
- Molecular Biology
- Microbiology
- Epidemiology
Context:
- Rickettsial infections, including spotted fever group (SFG) rickettsiosis and epidemic typhus, pose significant public health concerns.
- Accurate identification of Rickettsia species is crucial for diagnosis and epidemiological studies.
- Previous diagnostic methods may lack specificity for certain Rickettsia species.
Purpose:
- To develop and validate novel primer pairs for the Polymerase Chain Reaction (PCR) assay.
- To specifically detect and differentiate Rickettsia japonica from other Rickettsia species.
- To identify the etiological agent of SFG rickettsiosis in human and tick samples from Kanagawa prefecture.
Summary:
- Primer pairs R1/R2 were designed to amplify a 540-bp fragment from various SFG and epidemic typhus rickettsiae, including R. japonica. Specific primers Rj5/Rj10 amplified a 357-bp fragment exclusively from R. japonica and the Katayama strain.
- The Katayama strain was identified as R. japonica using primers Rj5/Rj10. Human blood samples from Kanagawa prefecture yielded both 537-bp and 357-bp fragments, indicating R. japonica as the causative agent of SFG rickettsiosis.
- Ixodes ovatus and Haemaphysalis flava ticks collected in Kanagawa prefecture also yielded both fragments, confirming them as vectors of R. japonica. The presence of SFG rickettsiae other than R. japonica was also suggested in these tick species.
Impact:
- This study provides a valuable molecular tool for the precise identification of Rickettsia japonica.
- The findings enhance our understanding of SFG rickettsiosis epidemiology in Japan, identifying key tick vectors.
- The developed primers can aid in rapid and accurate diagnosis, contributing to effective disease management and control strategies.
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