Rapid identification and quantification of Collinsella aerofaciens using PCR.
The number and incidence of Collinsella aerofaciens in the human intestine are the highest among Gram-positive non-spore-forming bacilli. Identification of this species is very difficult and requires considerable time. A PCR-based identification system using C. aerofaciens-specific primers is described. Using this PCR method, we identified 181 C. aerofaciens-like species isolated from human feces. These 181 strains were identified using the traditional method in past studies. Results of both methods matched. The direct detection method was performed using human feces samples from seven adults. Nested PCR was applied directly to the samples and all seven samples were positive. Quantification studies were performed using LightCycler¿trade mark omitted¿. The assay uses a double-stranded DNA dye to continuously monitor product formation and in a short time is able to quantify samples to 5 log units in concentration.
Kageyama
A
A
Japan Collection of Microorganisms, The Institute of Physical and Chemical Research (RIKEN), Wako-shi, Saitama, Japan. kageyama@jcm.riken.go.jp
Sakamoto
M
M
Benno
Y
Y
D039903:Actinobacteria / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D000328:Adult; D015169:Colony Count, Microbial; D004247:DNA / Q000378:metabolism; D017931:DNA Primers; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics; D004275:DNA, Ribosomal / Q000032:analysis / Q000235:genetics; D005243:Feces / Q000382:microbiology*; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012336:RNA, Ribosomal, 16S / Q000235:genetics*; D013045:Species Specificity
D017931:DNA Primers; D004269:DNA, Bacterial; D004275:DNA, Ribosomal; D012336:RNA, Ribosomal, 16S; D004247:DNA