PMID- 11390674
OWN - NLM
STAT- MEDLINE
DCOM- 20011101
LR  - 20200826
IS  - 1350-0872 (Print)
IS  - 1350-0872 (Linking)
VI  - 147
IP  - Pt 6
DP  - 2001 Jun
TI  - Visualization of Borrelia burgdorferi sensu lato by fluorescence in situ 
      hybridization (FISH) on whole-body sections of Ixodes ricinus ticks and gerbil 
      skin biopsies.
PG  - 1425-1436
LID - 10.1099/00221287-147-6-1425 [doi]
AB  - The objective of this study was to visualize borreliae directly in whole-body 
      sections of Ixodes ricinus by fluorescence in situ hybridization (FISH). Borrelia 
      afzelii mono-infected or Borrelia burgdorferi sensu stricto (ss)/B. afzelii 
      double-infected nymphs were fixed, embedded in cold polymerizing resin and 
      sectioned. The same sample processing was applied to skin biopsies taken from a 
      Mongolian gerbil after an infectious tick-bite. FISH was carried out using 
      16S-rRNA-directed, fluorescence-labelled oligonucleotide probes specific for the 
      genus Borrelia and specific within the group of Lyme borreliosis-associated 
      genospecies B. afzelii, B. burgdorferi ss, Borrelia garinii and Borrelia 
      valaisiana. Sensitivity and specificity of the newly designed probes were 
      evaluated using PCR, dot-blot hybridizations and FISH. Despite significant 
      autofluorescence of certain tick tissues (which allowed good histological 
      orientation within the sections), borreliae showing the typical spirochaetal 
      morphotype were clearly visible in five out of six putatively infected ticks. 
      These findings were confirmed by electron microscopy of ticks from the same 
      infected batch as used for FISH. Attempts to produce ticks infected by two 
      different Borrelia genospecies were not successful. FISH on whole-body sections 
      of resin-embedded ticks offers the possibility of visualizing and identifying 
      borreliae within tick tissues. This technique has great potential for the 
      investigation of the transmission of bacteria or other micro-organisms by 
      arthropod vectors. Furthermore, clear visualization of single spirochaetes 
      distributed along subcutaneous fat cell membranes in gerbil skin biopsies 
      suggests that FISH might also be suitable for the detection of borreliae in 
      clinical tissue specimens.
FAU - Hammer, Bettina
AU  - Hammer B
AD  - Institut fur Mikrobiologie und Hygiene, Universitatsklinikum Charite, 
      Humboldt-Universitat zu Berlin, Dorotheenstrabetae 96, D-10117 Berlin, Germany1.
FAU - Moter, Annette
AU  - Moter A
AD  - Institut fur Mikrobiologie und Hygiene, Universitatsklinikum Charite, 
      Humboldt-Universitat zu Berlin, Dorotheenstrabetae 96, D-10117 Berlin, Germany1.
FAU - Kahl, Olaf
AU  - Kahl O
AD  - Institut fur Biologie, Angewandte Zoologie/Okologie der Tiere, Freie Universitat 
      Berlin, Haderslebener Strabetae 9, D-12163 Berlin, Germany2.
FAU - Alberti, Gerd
AU  - Alberti G
AD  - Zoologisches Institut und Museum, Ernst-Moritz-Arndt-Universitat, 
      J.-S.-Bachstrabetae 11/12, D-17487 Greifswald, Germany3.
FAU - Gobel, Ulf B
AU  - Gobel UB
AD  - Institut fur Mikrobiologie und Hygiene, Universitatsklinikum Charite, 
      Humboldt-Universitat zu Berlin, Dorotheenstrabetae 96, D-10117 Berlin, Germany1.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Microbiology (Reading)
JT  - Microbiology (Reading, England)
JID - 9430468
RN  - 0 (DNA, Bacterial)
RN  - 0 (DNA, Ribosomal)
RN  - 0 (RNA, Ribosomal, 16S)
SB  - IM
MH  - Animals
MH  - Borrelia burgdorferi Group/growth & development/*ultrastructure
MH  - DNA, Bacterial/analysis
MH  - DNA, Ribosomal/analysis
MH  - Fluorescent Antibody Technique, Indirect
MH  - Gerbillinae
MH  - In Situ Hybridization, Fluorescence
MH  - Ixodes/*microbiology
MH  - Larva
MH  - Microscopy, Electron
MH  - Polymerase Chain Reaction
MH  - RNA, Ribosomal, 16S/analysis
MH  - Skin/*microbiology
EDAT- 2001/06/08 10:00
MHDA- 2001/11/03 10:01
CRDT- 2001/06/08 10:00
PHST- 2001/06/08 10:00 [pubmed]
PHST- 2001/11/03 10:01 [medline]
PHST- 2001/06/08 10:00 [entrez]
AID - 10.1099/00221287-147-6-1425 [doi]
PST - ppublish
SO  - Microbiology (Reading). 2001 Jun;147(Pt 6):1425-1436. doi: 
      10.1099/00221287-147-6-1425.