Development and use of a simple polymerase chain reaction assay to screen for Helicobacter spp. and H. hepaticus in intestinal and fecal samples from laboratory mice.
A simple and sensitive duplex polymerase chain reaction (PCR) assay was developed for use in detection of Helicobacter species and H. hepaticus in laboratory mice. Bacteria were extracted and concentrated from fecal pellets and intestinal segments by use of buoyant density centrifugation. To improve quality assurance, an internal control (mimic) for detection of false-negative reactions was included. In addition, cartridges (Capillette) pre-filled with PCR reagents, were used to minimize the hands-on time required, thus reducing the risk of contamination with previously amplified material. Laboratory mice from Swedish animal houses sent to the National Veterinary Institute for health monitoring were found to have high prevalence of H. hepaticus.
Fermér
Christian
C
National Veterinary Institute, SE-751 89 Uppsala, Sweden.
Lindberg
Anna V
AV
Feinstein
Ricardo E
RE
D000822:Animal Husbandry / Q000379:methods; D000818:Animals; D000830:Animals, Laboratory / Q000382:microbiology*; D002432:Cecum / Q000382:microbiology; D003106:Colon / Q000382:microbiology; D017931:DNA Primers / Q000737:chemistry; D004269:DNA, Bacterial / Q000032:analysis; D005188:False Negative Reactions; D005243:Feces / Q000382:microbiology; D005260:Female; D016998:Helicobacter / Q000235:genetics / Q000302:isolation & purification*; D016481:Helicobacter Infections / Q000175:diagnosis / Q000382:microbiology / Q000662:veterinary*; D008297:Male; D051379:Mice; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D012376:Rodent Diseases / Q000175:diagnosis / Q000378:metabolism*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004269:DNA, Bacterial