Detection of Rickettsia japonica in Haemaphysalis longicornis ticks by restriction fragment length polymorphism of PCR product

Abstract

PCR was applied to the detection of Rickettsia japonica, the causative agent of Oriental spotted fever (OSF), in ticks collected at two sites of the Muroto area on Shikoku Island, a major area in Japan where OSF is endemic. Primer pair Rr190.70p and Rr190.602n of the R. rickettsii 190-kDa antigen gene sequence of Regnery and others (R. L. Regnery, C. L. Spruill, and B. D. Plikaytis, J. Bacteriol. 173:1576-1589, 1991) primed the DNA extracted from Haemaphysalis longicornis ticks but not those extracted from Haemaphysalis formosensis, Haemaphysalis flava, Haemaphysalis hystricis, or Amblyomma testudinarium ticks. Digestion of the amplification product with the restriction endonucleases PstI and AluI produced the restriction fragment length polymorphism pattern specific to R. japonica. The HindIII and MspI digests gave restriction fragment length polymorphism patterns identical to those of the PCR product from R. japonica DNA. Hemolymph preparations of H. longicornis ticks were demonstrated to contain rod-shaped organisms that were detected by immunofluorescence with the monoclonal antibody specific to R. japonica species. The primer pair did not amplify the DNA of a laboratory colony of H. longicornis ticks originally collected at an area where OSF is not endemic. Our results provided evidence that H. longicornis ticks might be an arthropod reservoir for R. japonica and a vector of OSF.

Phylogenetic Analysis of Spotted Fever Group Rickettsiae Based on gltA, 17‐kDa, and rOmpA Genes Amplified by Nested PCR from Ticks in Japan

“…japonica has been isolated from Dermacentor taiwanensis and H. flava in the endemic areas of Japanese spotted fever (8). The rOmpA gene of R. japonica was detected from H. longicornis and H. formosenis by Uchida et al (40). In the present study, however, the genes of R. japonica were not detected by PCR from ticks even in the endemic areas of Japanese spotted fever.…”

Rickettsioses as paradigms of new or emerging infectious diseases

“…The disease is now known to be endemic in the southwestern part of Japan, where more than 100 cases have been described. The agent, R. japonica (292), and its tick vectors, Haemaphysalis longicornis (261) and Dermacentor taiwanensis (244), have now been characterized. The ticks Ixodes ovatus and H. flava have also been incriminated as vectors of this rickettsia.…”

<span style="font-family: 'Times New Roman','serif'; font-size: 12pt; mso-fareast-font-family: 'Malgun Gothic'; mso-fareast-theme-font: minor-fareast; mso-ansi-language: EN-US; mso-fareast-language: EN-US; mso-bidi-language: AR-SA;"><strong>Tick Surveillance in Four Southwestern Provinces of the Republic of Korea during 2013</strong></span>

“…Ticks belonging to the family Ixodidae, or hard ticks, are obligate blood-feeding ectoparasites of all classes of terrestrial vertebrates (mammals, birds, reptiles, and amphibians) and are vectors of emerging infectious diseases that pose veterinary and medical threats worldwide (Parola & Raoult 2001). Although tick-borne diseases were not reported in the Republic of Korea (ROK) until after 1973 (Chow 1973), ticks are now recognized as important vectors of a wide range of viral, bacterial, and protozoan pathogens in the ROK (Park et al 1992, Shim et al 1993, Uchida et al 1995, 2008, Choi, et al 2005, Kang et al 2013. As a result, tick-borne disease surveillance has taken on increasing importance as more pathogens affecting human health are discovered (Cho et al 1991, Kee et al 1994, Sachar 2000, Heo et al 2002, Jang et al 2004, including the presence in 2013 of a novel Bunyavirus, severe fever with thrombocytopenia syndrome virus (SFTSV) [also called Huaiyangshan virus (HYSV)], that resulted in 35 reported cases with 16 deaths (45.7% mortality) (Zhang et al 2012.…”