Real‐Time Quantitative Polymerase Chain Reaction Analysis of Patients With Refractory Chronic Periodontitis
Uložené v:
| Názov: | Real‐Time Quantitative Polymerase Chain Reaction Analysis of Patients With Refractory Chronic Periodontitis |
|---|---|
| Autori: | Simone M., Covani U., Barone A., Vittorio O., Curcio M., Barbuti S., Scatena F., FELLI, LAMBERTO, NICOLINI, CLAUDIO, MARCONCINI, SIMONE |
| Zdroj: | Journal of Periodontology, Vol. 82, No 7 (2011) pp. 1018-1024 |
| Informácie o vydavateľovi: | Wiley, 2011. |
| Rok vydania: | 2011 |
| Predmety: | Adult, Male, 0301 basic medicine, Polymerase Chain Reaction/methods, Bioinformatics, Genes, Periodontitis, Polymerase chain reaction, Gene Expression Regulation/genetics, Polymerase Chain Reaction, 03 medical and health sciences, Humans, Genetic Predisposition to Disease, GRB2 Adaptor Protein/genetics, Glyceraldehyde-3-Phosphate Dehydrogenases/genetics, Chronic Periodontitis/genetics, GRB2 Adaptor Protein, NF-kappa B p50 Subunit/genetics, Transcription Factor RelA/genetics, 0303 health sciences, Aggressive Periodontitis/genetics, Genetic Predisposition to Disease/genetics, Oncogene Protein v-cbl/genetics, Transcription Factor RelA, Computational Biology, Glyceraldehyde-3-Phosphate Dehydrogenases, NF-kappa B p50 Subunit, Middle Aged, Oncogene Protein v-cbl, 3. Good health, Aggressive Periodontitis, Gene Expression Regulation, Chronic Periodontitis, Disease Progression, Female |
| Popis: | Background: Periodontitis is a complex multifactorial 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 patients with 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/μL 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 groups were compared to the analysis of variance. A probability value Results: The present study agrees with the preliminary bioinformatics analysis. In our experiments, the association of pathology with the genes was statistically significant for GRB2 and CBL (P Conclusion: This article lends support to our preliminary hypothesis that assigned an important role in refractory aggressive periodontitis to leader genes. |
| Druh dokumentu: | Article |
| Popis súboru: | application/pdf |
| Jazyk: | English |
| ISSN: | 1943-3670 0022-3492 |
| DOI: | 10.1902/jop.2010.100312 |
| Prístupová URL adresa: | https://pubmed.ncbi.nlm.nih.gov/21189087 https://arpi.unipi.it/handle/11568/199757 https://core.ac.uk/display/80238072 https://archive-ouverte.unige.ch/unige:108486 https://pubmed.ncbi.nlm.nih.gov/21189087/ https://www.joponline.org/doi/pdf/10.1902/jop.2010.100312 https://aap.onlinelibrary.wiley.com/doi/full/10.1902/jop.2010.100312 https://archive-ouverte.unige.ch/unige:108486 https://doi.org/10.1902/jop.2010.100312 https://archive-ouverte.unige.ch/unige:108486 https://hdl.handle.net/11568/199757 https://doi.org/10.1902/jop.2010.100312 |
| Rights: | Wiley Online Library User Agreement |
| Prístupové číslo: | edsair.doi.dedup.....a63b70001e94a547f46d1b5b0244a6a6 |
| Databáza: | OpenAIRE |
Full Text Finder 
Nájsť tento článok vo Web of Science | Abstrakt: | Background: Periodontitis is a complex multifactorial 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 patients with 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/μL 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 groups were compared to the analysis of variance. A probability value Results: The present study agrees with the preliminary bioinformatics analysis. In our experiments, the association of pathology with the genes was statistically significant for GRB2 and CBL (P Conclusion: This article lends support to our preliminary hypothesis that assigned an important role in refractory aggressive periodontitis to leader genes. |
|---|---|
| ISSN: | 19433670 00223492 |
| DOI: | 10.1902/jop.2010.100312 |