Abstract
Background: Periodontitis is a complexmultifactorial disease and is typically polygenic in origin.Genes play a fundamental part in each biologic process forming complex networks of interactions. However, only some genes have a high number of interactions with other genes in the network and may, therefore, be considered to play an important role. In a preliminary bioinformatic analysis, five genes that showed a higher number of interactions were identified and termed leader genes. In the present study, we use real-time quantitative polymerase chain reaction (PCR) technology to evaluate the expression levels of leader genes in the leukocytes of 10 patientswith refractory chronic periodontitis and compare the expression levels with those of the same genes in 24 healthy patients. Methods: Blood was collected from 24 healthy human subjects and 10 patients with refractory chronic periodontitis and placed into heparinized blood collection tubes by personnel trained in phlebotomy using a sterile technique. Blood leukocyte cells were immediately lysed by using a kit for total RNA purification from human whole blood. Complementary DNA (cDNA) synthesis was obtained from total RNA and then real-time quantitative PCR was performed. PCR efficiencies were calculated with a relative standard curve derived from a five cDNA dilution series in triplicate that gave regression coefficients >0.98 and efficiencies >96%. The standard curves were obtained using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and growth factor receptor binding protein 2 (GRB2), casitas B-lineage lymphoma (CBL), nuclear factor-KB1 (NFKB1), and REL-A (gene for transcription factor p65) gene primers and amplified with 1.6, 8, 40, 200, and 1,000 ng/mL total cDNA. Curves obtained for each sample showed a linear relationship between RNA concentrations and the cycle threshold value of real-time quantitative PCR for all genes. Data were expressed as mean ± SE (SEM). The groupswere compared to the analysis of variance.Aprobability value
| Original language | English |
|---|---|
| Pages (from-to) | 1018-1024 |
| Number of pages | 7 |
| Journal | Journal of Periodontology |
| Volume | 82 |
| Issue number | 7 |
| DOIs | |
| Publication status | Published - Jul 2011 |
Fingerprint
Keywords
- Bioinformatics
- Genes
- Periodontitis
- Polymerase chain reaction
ASJC Scopus subject areas
- Periodontics
Cite this
Real-Time quantitative polymerase chain reaction analysis of patients with refractory chronic periodontitis. / Marconcini, Simone; Covani, Ugo; Barone, Antonio; Vittorio, Orazio; Curcio, Michele; Barbuti, Serena; Scatena, Fabrizio; Felli, Lamberto; Nicolini, Claudio.
In: Journal of Periodontology, Vol. 82, No. 7, 07.2011, p. 1018-1024.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Real-Time quantitative polymerase chain reaction analysis of patients with refractory chronic periodontitis
AU - Marconcini, Simone
AU - Covani, Ugo
AU - Barone, Antonio
AU - Vittorio, Orazio
AU - Curcio, Michele
AU - Barbuti, Serena
AU - Scatena, Fabrizio
AU - Felli, Lamberto
AU - Nicolini, Claudio
PY - 2011/7
Y1 - 2011/7
N2 - Background: Periodontitis is a complexmultifactorial disease and is typically polygenic in origin.Genes play a fundamental part in each biologic process forming complex networks of interactions. However, only some genes have a high number of interactions with other genes in the network and may, therefore, be considered to play an important role. In a preliminary bioinformatic analysis, five genes that showed a higher number of interactions were identified and termed leader genes. In the present study, we use real-time quantitative polymerase chain reaction (PCR) technology to evaluate the expression levels of leader genes in the leukocytes of 10 patientswith refractory chronic periodontitis and compare the expression levels with those of the same genes in 24 healthy patients. Methods: Blood was collected from 24 healthy human subjects and 10 patients with refractory chronic periodontitis and placed into heparinized blood collection tubes by personnel trained in phlebotomy using a sterile technique. Blood leukocyte cells were immediately lysed by using a kit for total RNA purification from human whole blood. Complementary DNA (cDNA) synthesis was obtained from total RNA and then real-time quantitative PCR was performed. PCR efficiencies were calculated with a relative standard curve derived from a five cDNA dilution series in triplicate that gave regression coefficients >0.98 and efficiencies >96%. The standard curves were obtained using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and growth factor receptor binding protein 2 (GRB2), casitas B-lineage lymphoma (CBL), nuclear factor-KB1 (NFKB1), and REL-A (gene for transcription factor p65) gene primers and amplified with 1.6, 8, 40, 200, and 1,000 ng/mL total cDNA. Curves obtained for each sample showed a linear relationship between RNA concentrations and the cycle threshold value of real-time quantitative PCR for all genes. Data were expressed as mean ± SE (SEM). The groupswere compared to the analysis of variance.Aprobability value
AB - Background: Periodontitis is a complexmultifactorial disease and is typically polygenic in origin.Genes play a fundamental part in each biologic process forming complex networks of interactions. However, only some genes have a high number of interactions with other genes in the network and may, therefore, be considered to play an important role. In a preliminary bioinformatic analysis, five genes that showed a higher number of interactions were identified and termed leader genes. In the present study, we use real-time quantitative polymerase chain reaction (PCR) technology to evaluate the expression levels of leader genes in the leukocytes of 10 patientswith refractory chronic periodontitis and compare the expression levels with those of the same genes in 24 healthy patients. Methods: Blood was collected from 24 healthy human subjects and 10 patients with refractory chronic periodontitis and placed into heparinized blood collection tubes by personnel trained in phlebotomy using a sterile technique. Blood leukocyte cells were immediately lysed by using a kit for total RNA purification from human whole blood. Complementary DNA (cDNA) synthesis was obtained from total RNA and then real-time quantitative PCR was performed. PCR efficiencies were calculated with a relative standard curve derived from a five cDNA dilution series in triplicate that gave regression coefficients >0.98 and efficiencies >96%. The standard curves were obtained using glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and growth factor receptor binding protein 2 (GRB2), casitas B-lineage lymphoma (CBL), nuclear factor-KB1 (NFKB1), and REL-A (gene for transcription factor p65) gene primers and amplified with 1.6, 8, 40, 200, and 1,000 ng/mL total cDNA. Curves obtained for each sample showed a linear relationship between RNA concentrations and the cycle threshold value of real-time quantitative PCR for all genes. Data were expressed as mean ± SE (SEM). The groupswere compared to the analysis of variance.Aprobability value
KW - Bioinformatics
KW - Genes
KW - Periodontitis
KW - Polymerase chain reaction
UR - http://www.scopus.com/inward/record.url?scp=79960523655&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=79960523655&partnerID=8YFLogxK
U2 - 10.1902/jop.2010.100312
DO - 10.1902/jop.2010.100312
M3 - Article
C2 - 21189087
AN - SCOPUS:79960523655
VL - 82
SP - 1018
EP - 1024
JO - Journal of Periodontology
JF - Journal of Periodontology
SN - 0022-3492
IS - 7
ER -