Article Figures & Data
Figures
- FIG. 1.
Representative gel electrophoresis of 16S rDNA type-specific PCR. Lanes 1 to 9, BH1 type-specific PCR; lanes 10 to 18, BH2 type-specific PCR. Lanes 1 and 10, JR1; lanes 2 and 11, ATCC 49793; lanes 3 and 12, ATCC 49882 (Houston-1); lanes 4 and 13, URLLY-8 (Marseille); lanes 5 and 14, R1073; lanes 6 and 15, NU4423; lanes 7 and 16, HC35; lanes 8 and 17, NU4428; lanes 9 and 18, negative (water). M, 0.07- to 12.2-kbp marker (Roche Diagnostics, Mannheim, Germany).
- FIG. 2.
(A) Representative HaeIII restriction fragment length polymorphism patterns of PCR-amplified 16S-23S ITS region. Lane 1, JR2; lane 2, JR3; lane 3, URLLY-8 (Marseille); lane 4, ATCC 49882 (Houston-1); lane 5, ATCC 49793; lane M, 0.07- to 12.2-kbp marker (Roche) (sizes are indicated on the right-hand margin); lane 6, HC63; lane 7, NU4428; lane 8, NU4714; lane 9, RMC1; lane 10, negative control (water). (B) 16S-23S rRNA ITS sequence of B. henselae Houston-1 (GenBank accession no. L35101 ). The tRNA, 16S, and 23S rRNA genes are underlined. The repeat region is boxed, and arrows denote base substitutions or insertions which vary from the Houston-1 sequence, as follows: 1, G-to-A substitution in HC35, HC54, HC60, HC62, HC69, NU4681, NU4714, JR2, BH2, R987, and R1073; 2, G-to-A substitution in HC54, HC69, NU4681, JR2, and BH2; 3, T-to-G substitution in HC35 and HC69; 4, C-to-T substitution in HC69; 5, C-to-T substitution in HC35, HC60, HC62, NU4428, NU4681, NU4714, RMC1, RMC8, RMC12, R987, and R1073; 6, A-to-G substitution in HC35, HC54, HC60, HC62, HC69, NU4428, NU4714, RMC1, RMC3, RMC8, RMC10, RMC12, R1073, R987, and JR2; 7, G-to-A substitution in HC60, HC62, NU4428, NU4714, RMC1, RMC8, RMC12, R987, and R1073. (C) 16S-23S ITS repeat region. There are two ATTGCTTCTAAAAAG repeats in HC54, HC69, RMC3, and JR2; three repeats in RMC10; and one repeat in BH2. The boxed region illustrates the previously described repeat sequence in B. henselae SA-2 (14).
- FIG. 3.
AP-PCR representative gel. Lane 1, ATCC 49882 (Houston-1); lane 2, JR2; lane 3, RMC1; lane 4, BH2; lane 5, RMC8; lane 6, URLLY-8 (Marseille); lane 7, HC63; lane 8, HC77; lane 9, BH4; lane 10, NU4428; M, pGEM marker (Promega; molecular size in base pairs is given along the right border).
- FIG. 4.
ERIC-PCR. (A) Representative gel electrophoresis of ERIC-PCR. Lanes 1 to 3, pattern E4H (strains ATCC 49882 [Houston-1] and NU4714); lanes 4 and 5, pattern E4M (strains BH4 and Marseille); lanes 6 and 7, pattern E2 (strains HC61 and NU4428); lanes 8 and 9, patterns E3a and -b, respectively (strains HC35 and HC60); lanes 10 and 11, pattern E1 (strains HC62 and RMC1); lanes 12 and 13, pattern E5 (strains RMC3 and RMC7); lanes 14 and 15, pattern E6 (strain RMC2 duplicate); lanes 16 and 17, pattern E7 (strains RMC11 and RMC4); lanes M1 and M2, molecular size markers. (B) Dendrogram. B. quintana and B. vinsonii patterns are quite distinct from all others (not shown in panel A) and are included in a merged reconstructed dendrogram. The dendrogram reflects the similarity of distribution of ERIC priming sites and is derived from gels (including that shown in panel A), all of which included B. henselae and the two molecular size markers and were analyzed identically.
- FIG. 5.
EagI-HhaI IRS-PCR. Lane 1, negative control; lane 2, NU4713; lane 3, HC61; lane 4, BH2; lane 5, R1073; lane 6, Marseille; lane 7, ATCC 49882 (Houston-1); lane 8, NU4714; lane 9, ATCC 49793; lane 10, HC35; lane 11, HC60; lane M, pGEM marker (Promega) (molecular sizes are given along the right border).
Tables
- TABLE 1.
B. henselae isolates studieda
Isolate(s) Source 16S type gltA type AP type ERIC type IRS type Feline isolates (n = 31) NU4695 Blood I H H E4H 1 RMC10† Blood I H H E4H 1 HC54 Blood I H H E4H 2 HC69 Blood I H M E4H 2 HC62† Blood I H H E1 2 RMC3* Blood I H H E5 2 NU4428† Blood I H M E2 2 HC71 Blood I M H E2 2 NU4714*† Blood II H H E4H 1 HC35*, HC55 Blood II H H E3a 1 RMC8* Blood II H Ha E3a 2 NU4681* Blood II H H E3a 1 HC60† Blood II H H E3b 1 RMC1* Blood II H H E1 2 RMC12 Blood II H M E3a 1 NU4423† Blood II M H E4H 2 RMC11 Blood II M M E7 1 RMC4 Blood II M M E7 2 HC41, HC48†, HC61, HC63†, HC77 Blood II M M E2 2 RMC2† Blood II M M E6 2 RMC5 Blood II M M E1 2 RMC6†, RMC9† Blood II M M E5 2 RMC7, Berlin-2 Blood II M M E5 2 NU4713 Blood II M H E2 2 Human isolates (n = 28) BH2 CSD I H H E4H 1 ATCC 49793/NZRm3492 (s4) BA I H H E4H 1 ATCC 49882 (Houston-1)†, Berlin-1 BA I H H E4H 2 BH3(s4), BH5s2*, R987, JR1, JR2, JR3, JR5, JR6, JR7, JR8, JR9, JR12, JR13, JR14, JR15, JR17, JR18, JR19, JR20, JR22, JR23 CSD I H H E4H 2 R1073† CSD II H H E4H 2 BH4 CSD II M M E4M 2 URLLY-8 (Marseille)† BA II M M E4M 2 ↵ a Symbols: *, Isolate embeds into solid agar and displays ratchet motility at this passage number; †, 16S type was determined by direct sequencing. s2, second passage after initial isolation; (s4), third to fifth passage after initial isolation. If not specified, RMC-prefix isolates (this work) were studied at passage numbers 2 to 3; HC-prefix isolates were a gift of Tom Gottlieb (8) and were studied at passage numbers 4 to 6; NU-prefix isolates (15) were studied at passage numbers 3 to 5; R- and JR-prefix strains (this work and Fournier et al. [12a]) were all studied at passage numbers 4 to 7. Other isolates were isolated in our laboratory or obtained from the following: Mardjan Arvand, Hygiene-Institute, Heidelberg, Germany (2) (Berlin-1 and -2); R. Bunter, Goulburn Valley Base Hospital, Goulburn, NSW, Australia (BH2); the New Zealand quality control strain NZRm3492 is thought to be a lab-passaged derivative of the widely used strain ATCC 49793, and they are inseparable by all typing methods; Jock Harkness, Royal North Shore Hospital, Sydney, Australia [BH3(s4)]; T.H.E. Pathology, Fairfield Heights, NSW, Sydney, Australia [BH5(S2)]; and D. Raoult, Unite des Rickettsies, Marseille, France (9) (URLLY-8 [Marseille]). Strains with the ATCC prefix were obtained from the American Type Culture Collection, Manassas, Va. 16S-23S regions were sequenced in all feline strains, in all Sydney and imported human strains, and in JR2.
