title: Comparison of two laboratory extraction techniques for the detection of Epstein-Barr virus in the saliva of nasopharyngeal carcinoma patients.
abstract: OBJECTIVE
  The aim of the present study was to compare the effectiveness of DNA extraction using an extraction kit against the standard boiling technique for the detection of Epstein-Barr virus (EBV) DNA in nasopharyngeal carcinoma (NPC) patients.
  
  
  METHODS
  Stimulated whole saliva samples from newly-diagnosed NPC patients were collected. EBV DNA was extracted by both techniques (n = 23) followed by quantitative real-time polymerase chain reaction (PCR) using the primer/probe set for BALF5.
  
  
  RESULTS
  The results of the quantitative real-time PCR were reproducible in both groups. The two techniques were moderately correlated (r = 0.67, P < 0.05), and the degree of agreement was good. However, the mean EBV DNA level in the boiling group (3.02 ± 8.67 × 10(6) copies/μL) was significantly higher than the extraction kit group (1.15 ± 2.66 × 10(6) copies/μL) (P < 0.05). The EBV DNA level was higher in patients at an advanced overall stage (P = 0.05).
  
  
  CONCLUSIONS
  The results of the present study showed that the performance of the extraction kit was not superior to the simple boiling technique for the detection of salivary EBV DNA in NPC patients using real-time PCR. The salivary EBV DNA level in patients at an advanced overall stage appeared to be higher than in patients at an early stage.
authors.affiliation: Discipline in Oral Rehabilitation, Faculty of Dentistry, The University of Hong Kong, Hong Kong, China.
authors.forename: Junwen
authors.initials: J
authors.lastname: Shan
authors.forename: Edmond H N
authors.initials: EH
authors.lastname: Pow
authors.forename: Peter C S
authors.initials: PC
authors.lastname: Tsang
authors.forename: Ranawaka A P M
authors.initials: RA
authors.lastname: Perera
authors.forename: Dora L W
authors.initials: DL
authors.lastname: Kwong
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D002277:Carcinoma / Q000821:virology*; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D004268:DNA-Binding Proteins / Q000032:analysis; D004259:DNA-Directed DNA Polymerase / Q000032:analysis; D020031:Epstein-Barr Virus Infections / Q000821:virology; D005260:Female; D004854:Herpesvirus 4, Human / Q000235:genetics / Q000302:isolation & purification*; D006801:Humans; D008297:Male; D008875:Middle Aged; D009303:Nasopharyngeal Neoplasms / Q000821:virology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods; D012463:Saliva / Q000821:virology*; D014764:Viral Proteins / Q000032:analysis; D014776:Virus Cultivation
chemical_list: D017931:DNA Primers; D004279:DNA, Viral; D004268:DNA-Binding Proteins; D014764:Viral Proteins; C083202:BALF5 protein, Epstein-barr virus; D004259:DNA-Directed DNA Polymerase
keywords: DNA; Epstein-Barr virus; nasopharyngeal carcinoma; polymerase chain reaction; saliva