[Amplification and restriction fragment length polymorphism analysis on the complete sequence of Sta56 gene of Orientia tsutsugamushi isolated from Shandong area].
OBJECTIVE
To analyze the genetic differences of Orientia tsutsugamushi (Ot) Sta56 gene between Shandong isolates and other strains deposited in GenBank.


METHODS
PCR and restriction fragment length polymorphism (RFLP) were used to amplify the complete sequence of Ot-Sta56 gene. RFLP profiles of Ot were predicted by a computer program according to their complete sequences of Ot-Sta56 gene. PCR amplicon from XDM2 strain was sequenced and analyzed by Clustal X (1.8) and PHYLIP software.


RESULTS
The complete sequences (about 1.6 kbp) of Ot-Sta56 gene were amplified from B16 strain (isolated from patients), FXS2 strain (isolated from A. agrarius) and XDM2 strain. Four species of restriction endonucleases (Hha I, Hinf I, Hae III, Pst I) were used to digest the PCR amplicons from the 3 isolates. When comparing with the RFLP profiles of prototype Ot, the RFLP profiles of PCR amplicons from the 3 isolates were similar to those of Japan Kawasaki strain, but were quite different from the international reference strains Gilliam, Karp, Kato. Results from DNA sequence analysis showed that the complete sequence of Ot-Sta56 gene homology to Japan Kawasaki strain of XDM2 strain was 97%, and deduced amino acid sequence was 92%.


CONCLUSIONS
Data from the complete sequence of Sta56 gene indicated that the genotypes of Ot isolates in Shandong province were similar, but with distinction from the Kawasaki strain.
Liu
Yun-Xi
YX
General Hospital of PLA, Beijing 100853, China.
Zhang
Qian
Q
Zhao
Zhong-Tang
ZT
Yang
Zhan-Qing
ZQ
Yang
Li-Ping
LP
Zhang
Pan-He
PH
Yang
Hong
H
Yuan
Yun-E
YE
Wei
Hua
H
Suo
Ji-Jiang
JJ
Xing
Yu-Bin
YB
Jia
Ning
N
Gao
Yan
Y
Cao
Wu-Chun
WC
D054458:Amplified Fragment Length Polymorphism Analysis; D000942:Antigens, Bacterial / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics*; D015373:Bacterial Typing Techniques / Q000145:classification; D004269:DNA, Bacterial / Q000235:genetics; D005798:Genes, Bacterial; D008565:Membrane Proteins / Q000235:genetics*; D012285:Orientia tsutsugamushi / Q000235:genetics* / Q000302:isolation & purification; D012150:Polymorphism, Restriction Fragment Length; D017422:Sequence Analysis, DNA
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins