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Journal of Clinical Microbiology, September 2002, p. 3286-3290, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3286-3290.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Simultaneous Detection of Anaplasma marginale and a New Ehrlichia Species Closely Related to Ehrlichia chaffeensis by Sequence Analyses of 16S Ribosomal DNA in Boophilus microplus Ticks from Tibet

Bohai Wen,1* Rui Jian,2 Youzhi Zhang,3 and Rong Chen2

Department of Microbiology, Beijing Institute of Microbiology and Epidemiology, Beijing 100071,1 Department of Microbiology, Third Medical University, Chongqing,2 Chengdu Institute of Epidemiology, Chengdu, China3

Received 21 December 2001/ Returned for modification 18 May 2002/ Accepted 20 June 2002

To identify ehrlichial agents in Boophilus microplus ticks, DNA samples of B. microplus collected from the Tibet Autonomous Region and Sichuan Province of China were screened by a nested PCR. Sixteen of 43 (37%) DNA samples of B. microplus from Tibet were positive in nested PCR analysis. All 27 samples from Sichuan were negative. The screen identified two ehrlichial agents based on different 16S rRNA genes that were found after amplifying and sequencing the 5'-end fragments of the 16S rRNA genes. One sequence was identical to that of the gene of Anaplasma marginale, an etiological agent of animal anaplasmosis. The other sequence was most similar to that of the gene of Ehrlichia chaffeensis, an etiological agent of human monocytic ehrlichiosis. The sequence of 1,501 bases from the novel ehrlichial agent was obtained and showed the greatest levels of sequence similarity (97 to 98%) to 16S rRNA gene sequences of the members of the E. canis group of the genus Ehrlichia. Sequence comparison of the 16S rRNA gene with the members of the genus Ehrlichia reveals that the novel ehrlichial agent detected in B. microplus ticks is a new species of the genus Ehrlichia and is most closely related to E. chaffeensis.

* Corresponding author. Mailing address: Beijing Institute of Microbiology and Epidemiology, 20 Dong-Da-Jie St., Fengtai District, Beijing 100071, People's Republic of China. Phone: (086) 10-66948525. Fax: (086) 10-63815273. E-mail: bohaiwen{at}sohu.com.

Journal of Clinical Microbiology, September 2002, p. 3286-3290, Vol. 40, No. 9
0095-1137/02/$04.00+0     DOI: 10.1128/JCM.40.9.3286-3290.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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