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Journal of Clinical Microbiology, July 2007, p. 2138-2143, Vol. 45, No. 7
0095-1137/07/$08.00+0     doi:10.1128/JCM.00478-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Isolation and Propagation of the Ap-Variant 1 Strain of Anaplasma phagocytophilum in a Tick Cell Line{triangledown}

Robert F. Massung,1* Michael L. Levin,1 Ulrike G. Munderloh,2 David J. Silverman,3 Meghan J. Lynch,2 Jariyanart K. Gaywee,4 and Timothy J. Kurtti2

Viral and Rickettsial Zoonoses Branch, Centers for Disease Control and Prevention, Atlanta, Georgia 30333,1 Department of Entomology, University of Minnesota, Saint Paul, Minnesota 55108,2 School of Medicine, University of Maryland-Baltimore, Baltimore, Maryland 21201,3 Armed Forces Research Institute of Medical Sciences, Microbiology, Rajthawee, Bangkok, Thailand4

Received 2 March 2007/ Returned for modification 6 April 2007/ Accepted 24 April 2007

The first tissue culture isolates of the unique Anaplasma phagocytophilum strain, Ap-Variant 1, were obtained in the Ixodes scapularis tick-derived cell line ISE6. Two isolates were from goat blood samples: one from a goat infected with I. scapularis ticks from Rhode Island and a second from a goat infected by serial passage of blood from the first infected goat. Eight isolates were made directly from I. scapularis ticks collected from white-tailed deer in Minnesota and represent the first isolations of an Anaplasma species directly from ticks. Each of the 10 isolates had a 16S rRNA gene sequence identical to that previously described for Ap-Variant 1, but differences within the ank gene were found that suggest natural variation. Prevalence of Anaplasma in the Minnesota ticks was 63.9%; 23 of 36 ticks tested by PCR were positive. Six of the tick-derived isolates were obtained from a set of 18 PCR-positive ticks, for a 33.3% isolation success rate. The conservation of host tropism among the Rhode Island and Minnesota isolates of Ap-Variant 1 was examined by use of experimental infections of mice and a goat. A Minnesota tick-derived isolate (MN-61-2) was used to inoculate naïve animals, and this isolate was able to infect a goat but unable to infect each of five mice, confirming that the Minnesota isolates have the same host tropism as Ap-Variant 1 from the northeastern United States. Light and electron microscopy of the Ap-Variant 1 isolate MN-61-2 in ISE6 cells showed cytoplasmic inclusions characteristic of A. phagocytophilum with pleomorphic bacteria in membrane-bound vacuoles and both electron-dense and electron-lucent forms.

* Corresponding author. Mailing address: Centers for Disease Control and Prevention, 1600 Clifton Rd., MS G-13, Atlanta, GA 30333. Phone: (404) 639-1082. Fax: (404) 639-4436. E-mail: rfm2{at}cdc.gov

{triangledown} Published ahead of print on 2 May 2007.

Journal of Clinical Microbiology, July 2007, p. 2138-2143, Vol. 45, No. 7
0095-1137/07/$08.00+0     doi:10.1128/JCM.00478-07
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Krause, P. J., Wormser, G. P. (2008). Nosocomial Transmission of Human Granulocytic Anaplasmosis?. JAMA 300: 2308-2309 [Full Text]  


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