Objective Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007-2008, 28 human SFGR isolates and 34 rat SFGR isolates (including 15 isolates from Rattus fulvescens, 5 isolates from R. edwardsi, 7 isolates from Callosciurus erythraeus roberti, and 7 isolates from Dremomys rufigenis) were gathered by L929 cell culture. Previous research indicated that these isolates shared identical sequences of 16S rRNA,gltA, ompA, groEL, and 17×103 protein gene with each other, and were of the same serophenotype. This project aims to study the 62 SFGR isolates from fever patients and wild rodents in Chengmai county of Hainan province, China by multispacer typing (MST).Methods Nine pairs of primers which had been previously reported in MST of SFGR were used in amplification and sequencing of the 62 isolates in this study. Sequences were subjected to Blast analysis after data correction. Analysis of single nucleotide polymorphisms (SNPs) was performed with Megalign. Six spaced sequences that were successfully sequenced from each isolate were spliced and applied in the construction of phylogenetic tree. Results There were great variations among the 62 isolates. A total of 48 mutant genotypes were identified. The predominant genotype, h3, accounted for 21.7% (13/60) of all the isolates tested, and the remaining 47 genotypes were all unique, each accounting for 1.6% (1/60) of all isolates. Phylogenetic analysis showed that all these 48 genotypes were classified in the same cluster. Among all reference strains included in this study, R. heilongjiangensiswas genetically closest to these isolates. Thymine-to-cytosine transitions had the highest percentage (52/114, 45.6%) among all SNPs, followed by adenine-to-guanine transitions (37/114, 32.5%).Conclusion The non-coding spaced sequences of SFGR prevailing in Chengmai county of Hainan province are highly heterologous. The selected primers for MST of SFGR are capable of detecting rickettsial pathogens. MST can be used in the rapid and effective tracing of endemic strains in outbreak of rickettsiae.
CHENG Xue-qin, JIN Yu-ming, LAO Shi-jun, HUANG Chang-he, HUANG Fang, JIA Peng-ben, ZHANG Li-juan
. Multispacer typing of spotted fever group rickettsiae isolated from Chengmai county of Hainan province, China[J]. Chinese Journal of Vector Biology and Control, 2014
, 25(4)
: 289
-293
.
DOI: 10.11853/j.issn.1003.4692.2014.04.001
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