| Yazarlar |
Ersin Karataylı
Ankara Üniversitesi, Türkiye |
Prof. Dr. Yasemin ÇELİK ALTUNOĞLU
Kastamonu Üniversitesi, Türkiye |
|
Senem Ceren Özen Karataylı
Ankara Üniversitesi, Türkiye |
|
Süleyman Cihan Yurtaydın
Ankara Üniversitesi, Türkiye |
|
Abdurrahman Mithat Bozdayı
Ankara Üniversitesi, Türkiye |
| Özet |
| Internal controls (ICs), are the main components of any real-time PCR based amplification methods, which are co-purified and co-amplified with the actual target. The existence of free circulating nucleic acids in plasma and serum (CNAPS) has been known for many years. The aim of this study was to verify whether CNAPS can be used as ICs in real-time PCR based detection and quantification of DNA or RNA targets in plasma and serum samples. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) as a housekeeping gene, was chosen at random as CNAPS to serve as an intrinsic internal control in two different real-time PCR based quantification models in plasma and serum. Viral loads of hepatitis B virus (HBV) DNA and hepatitis delta virus (HDV) RNA were quantified as actual targets in parallel to GAPDH as IC in a total of 519 serum or plasma samples including 21 healthy controls, 202 positive chronic hepatitis delta patients, 37 chronic hepatitis C patients, 168 chronic hepatitis B patients, 52 patients with hepatocellular carcinoma, and 39 patients with non-alcoholic steatohepatitis/non-alcoholic fatty liver disease. GAPDH levels did not show significant variance in different patient groups and yielded positive signals in all 519 patients with persistent cycle threshold (CT) values 27.85±1.57 (mean±standard deviation (SD)). Reproducibility of the GAPDH amplification in HDV RNA and HBV DNA quantifications was shown with a SD value of CT ranging from 0.42 to 2.14 (mean SD; 1.18) and 0.24 to 1.75 (mean SD; 1.03), respectively. In conclusion, the freely circulating nucleic acids can clearly be used as internal controls for real-time PCR based detection and quantification of any RNA and mainly DNA targets (pathogens) in serum or plasma and this simply excludes the compulsory external addition of any IC molecules into the reaction. © 2014 Elsevier B.V. |
| Anahtar Kelimeler |
| Real-time PCR,Internal control,Circulating nucleic acids in serum or plasma,GAPDH |
| Makale Türü | Özgün Makale |
| Makale Alt Türü | SSCI, AHCI, SCI, SCI-Exp dergilerinde yayımlanan tam makale |
| Dergi Adı | Journal of Virological Methods |
| Dergi ISSN | 0166-0934 |
| Dergi Tarandığı Indeksler | SCI-Expanded |
| Makale Dili | İngilizce |
| Basım Tarihi | 07-2014 |
| Cilt No | 207 |
| Sayfalar | 133 / 137 |
| Doi Numarası | 10.1016/j.jviromet.2014.07.008 |
| Makale Linki | http://linkinghub.elsevier.com/retrieve/pii/S016609341400264X |
| Atıf Sayıları | |
| WoS | 9 |
| SCOPUS | 9 |
