FISH-Bact 2.0

DNA based identification of bacteria pathogenic to fish + IPNV 

  • Sensitivity
    short amplicon length (< 130-230 bp) for increased sensitivity with a broad spectrum of starting materials
  • Specificity
    8 bacterial species detected with one amplification distinguishes V. ordalii fom other Vibrio species (2cd amplification, optional)
    Including detection of Infectious Pancreatic Necrosis Virus (IPNV)
  • Fast Protocol
    optimized 45 minutes protocol including analysis (post amplification)
  • Simple & Robust
    Non-fluorescent detection for easy read-out, minimal lab instrumentation required

Product Information

Assay Principle

Biotinylated PCR products are generated from sample material containing extracted bacterial DNA or viral RNA. The labeled amplicons are hybridized to species specific capture probes immobilized on the LCD-Chip surface. Following a short wash routine, comprising high stringency, visualization of bound amplicons is mediated by an enzyme-substrate cascade. Each LCD-Chip contains eight identical micro arrays separated in small reaction chambers. Each array can be addressed individually, allowing the parallel analysis of eight samples on one chip.

 

Target Region & Sensitivity

The targets amplified by the supplied PCR* primer mixes are well conserved sequences and the probes spotted on the array have been thoroughly evaluated for their specificity. Bacteria: Two primer sets for PCR amplification are provided. A ‘broad’ mix targeting the 16S rRNA gene (~200 bp) and the Vibrio mix mix targeting a second 16S region to distinguish V. ordalii from other Vibrio species (~130 bp).

IPNV: A complex primer mixture allows to amplify a ~230 bp fragment of Infectious Pancreatic Necrosis Virus cDNA from almost all known strains / varieties .

 

Capture Probes

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Fig.1:

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Parallel analysis of eight samples on one LCD-Chip. The array field spacing is compatible with standard multi pipettes.

Fig.2 :

array-fish
A) Aeromonas salmonicida B) Flavobacterium psychrophilum
C) Renibacterium salmoninarum D) Aero. salmonicida + Vibrio sp.
E) Vibrio ordalii (2cd PCR) F) IPNV (3rd PCR)

* The Polymerase Chain Reaction (PCR) process is covered by U.S. patents owned by Hoffmann-La Roche, Inc. and patents owned by F. Hoffmann-La Roche Ltd. No license under these patents to use the PCR process is conveyed expressly or by implication to the purchaser by the purchase of this product.

 

Software

SlideReader Vers. 7.01

• Fully automated image analysis
• comprehensive data reports in HTML format
• fast & intuitive navigation
• multiple user accounts

 

Instrumentation

CHIP-Scanner PF2700

• Transmission light scanner for LCD-Arrays,

• 10 µm resolution

• 16 bit grey scale TIFF images

CHIP Spin FVL2400

• Bench Top mini centrifuge
• 2400 rpm, constant
• Adaptor for LCD-Arrays
• incl. 3 rotors

Order Information Cat. No.
FISH-Bact 1.5 LCD-Array Kit, 48 Tests C-120-06
CHIP-Scanner PF2700 HS-500-01
CHIP-Spin FVL2400 HS-300-01
SlideReader Software Vers. 7.00.01 HS-200-01

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