J Bacteriol Virol.  2008 Dec;38(4):221-226. 10.4167/jbv.2008.38.4.221.

Rapid Identification of Rickettsiae using the Real-Time PCR

• Park HS
• Lee JH
• Jin KH
• Jang WJ
• Park KH
• Kook YH
• Lee SH

• KMID: 1483962
• DOI: http://doi.org/10.4167/jbv.2008.38.4.221

Abstract

In this study, new real-time PCR method based on the groEL gene was developed and investigated. Four spotted fever group (SFG) strains, four typhus group (TG) strains, and four scrub typhus group (STG) strains were easily differentiated as a distinct entity. This PCR assay was applied to detect Rickettsia DNA from 100 ticks. Twelve Haemaphysalis longicornis ticks were found positive and identified as spotted fever group Rickettsia. This real-time PCR method could simultaneously perform the rapid identification of rickettsiae and the differential diagnosis of SFG, TG, and STG in a single reaction.

Keyword

groEL gene; Rickettsia; Real-time PCR

MeSH Terms

Diagnosis, Differential
DNA
Fever
Polymerase Chain Reaction
Real-Time Polymerase Chain Reaction
Rickettsia
Scrub Typhus
Ticks
Typhus, Epidemic Louse-Borne
DNA

Figure

• 

  Figure 1. Amplification of groEL DNA from rickettsial strains using the real-time PCR. Lanes; SFG (R japonica, R. conorii, R. sibirica, R akari), : TG (R. typhi, R. typhi 87~91, R. typhi 87~100, R prowazekii), STG (O. tsutsugamushi Karp, O. tsutsugamushi Kato, O. tsutsugamushi Boryong, O. tsutsugamushi Gilliam), : DW

• 

  Figure 2. Amplification of groEL DNA from rickettsial strains and tick DNA using the real-time PCR. Lanes; : SFG (R. japonica, R. conorii, R sibirica, R akari), : TG (R. typhi, R. typhi 87~91, R. typhi 87~100, R. prowazekii), : STG (O. tsutsugamushi Karp, O. tsutsugamushi Kato, O. tsutsugamushi Boryong, O. tsutsugamushi Gilliam), ————: tick (Tick no. 22, 28, 33, 36, 48, 52, 55, 61, 65, 68, 77, 92) : DW

• 

  Figure 3. The sensitivity of real-time PCR using the R. typhi plasmid. Lanes; (A) R. typhi 106, (B) R. typhi 105, (C) R. typhi 104, (D) R. typhi 103, (E) R. typhi 102, (F) R. typhi 101, (G) R. typhi 1, (H) Negative control.

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