Simple duplex fecal PCR assay that allows identification of false-negative results in Helicobacter sp.-infected mice.
We designed a simple and sensitive duplex polymerase chain reaction (PCR) assay for detection of false-negative results during routine Helicobacter sp. feces analysis. We took advantage of the various Lactobacillus species that form part of the normal intestinal flora of laboratory rodents to improve our PCR diagnostic assays. Using this one-step PCR assay, we were able to rule out false-negative results without the need of adding internal standard molecules. This is an important quality control for PCR diagnostic tests, since the presence of inhibitors in feces can prevent detection of Helicobacter infections using PCR analysis. Use of this Lactobacillus group-specific PCR assay can be extended to other feces tests used in mouse quality-control programs.
Bourgade
Franck
F
Animalerie Centrale, Institut Pasteur, 25 rue du Docteur Roux, 75724 Paris, France.
Montagutelli
Xavier
X
Bigbee
Cassie
C
Weiss
April
A
Rigottier-Gois
Lionel
L
Conti
Claudio J
CJ
Benavides
Fernando
F
D000818:Animals; D004269:DNA, Bacterial / Q000032:analysis*; D005188:False Negative Reactions; D005243:Feces / Q000382:microbiology*; D016998:Helicobacter / Q000235:genetics / Q000302:isolation & purification*; D007756:Laboratory Animal Science / Q000379:methods* / Q000592:standards; D048191:Lactiplantibacillus plantarum / Q000235:genetics; D051379:Mice; D008815:Mice, Inbred Strains; D016133:Polymerase Chain Reaction / Q000379:methods / Q000592:standards / Q000662:veterinary*; D011786:Quality Control; D015203:Reproducibility of Results
D004269:DNA, Bacterial