Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Pivot Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Target Concepts:
Gene/Protein
Disease
Symptom
Drug
Enzyme
Compound
Query: UMLS:C0031099 (
periodontitis
)
12,489
document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)
Two pairs of species-specific PCR primers targeting the housekeeping
groEL
gene, Spa146f-Spa525r and Spa93f-Spa525r, were designed to quantify human oral and fecal
Streptococcus parasanguinis
.
Blast
analysis against reference sequences of NCBI nucleotide collection database and the Chaperonin Sequence Database showed the forward primers Spa146f and Spa93f 100% matched only with
S. parasanguinis
, and the
in silico
Simulated PCR algorithm showed both primer pairs hit only
S. parasanguinis groEL
gene in Chaperonin Sequence Database. The two primer pairs were respectively used to perform PCR with saliva DNA of each of 6 human subjects, and the amplicons of individual PCR reactions were cloned. The phylogenetic analysis showed cloned sequences were all affiliated to
S. parasanguinis
, which further validates the specificity of two primer pairs, and that individual subjects harbored multiple genotypes of
S. parasanguinis
in saliva. By spiking
S. parasanguinis
into human fecal samples, we found the quantification limit of quantitative real-time PCR (qPCR) assays for both primer pairs was 5-6 log
10
groEL
copies/g feces. Human fecal
S. parasanguinis
amounts quantified with qPCR using each of the two primer pairs correlated well with those determined with metagenomic sequencing. qPCR with either primer pair showed
periodontitis
patients had significantly lower level of saliva
S. parasanguinis
than healthy people. In both feces and saliva, the
S. parasanguinis
abundances quantified with two primer pairs exhibited strong and significant correlation. Our results show that the two
S. parasanguinis
-specific primer pairs can be used to quantify and profile human saliva and fecal
S. parasanguinis
.
...
PMID:Quantification of Human Oral and Fecal
Streptococcus parasanguinis
by Use of Quantitative Real-Time PCR Targeting the
groEL
Gene. 3192 Oct 79
