Establishment of rapid diagnostic methods for rickettsial infections using loop-mediated isothermal amplification (LAMP)

2015 Fiscal Year Final Research Report

• Project/Area Number: 25860828
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Infectious disease medicine
• Research Institution: Shimane University
• Principal Investigator: NIIHARA HIROYUKI 島根大学, 医学部, 講師 (60362935)
• Co-Investigator(Renkei-kenkyūsha): KOHNO Kunie 島根大学, 医学部皮膚科学講座, 特別協力研究員 (20432619)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Keywords: 日本紅斑熱 / ツツガムシ病 / 病原体遺伝子検査 / LAMP / Real-ｔime PCR / 2段階PCR / リケッチア症

Outline of Final Research Achievements

In the present study, we propose a diagnostic flow towards the definite diagnosis of a rickettsial infection from the point of clinical importance. The first priority is an administration of antibiotic medication for the suspected cases. Next is to refer the diagnostic aids such as loop-mediated isothermal amplification (LAMP) and real-time PCR in an expeditious way. At the same time, 2-step PCR including nested PCR should be performed in order to catch up the exceptional cases such as Shimokoshi type of Orientia tsutsugamushi or unknown type of rickettsia. For definite diagnosis, the most precious way is an identification of pathogenic gene using sequencing analysis and phylogenetic tree analysis following to the nested PCR. In addition, we developed the new detection method for the rapid detection of Rickettsia japonica using LAMP method. We also confirmed of its utility from the specimen of definite cases with rickettsial infection.

Free Research Field

皮膚科学、感染症、薬疹、下肢静脈瘤