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Conference Paper: A comparison of laboratory extraction techniques for the detection of Epstein-Barr virus in saliva of nasopharyngeal carcinoma patients

TitleA comparison of laboratory extraction techniques for the detection of Epstein-Barr virus in saliva of nasopharyngeal carcinoma patients
Authors
Issue Date2009
PublisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/oraloncology
Citation
The 2nd World Congress of the International Academy of Oral Oncology "Oral Cancer: Imagine the Future", Toronto, Canada, 8-11 July 2009. In Oral Oncology Supplement, 2009, v. 3 n. 1, p. 200 Abstract no. P2.120 How to Cite?
AbstractNasopharyngeal carcinoma (NPC) is common in southern Chinese. It is closely associated with Epstein-Barr virus (EBV). Recent studies suggest that tumor burden is proportional to the level of EBV DNA in blood of NPC patients and that rapid blood testing can be used to guide therapeutic intervention. Instead of collecting invasive blood samples, saliva may be a viable alternative. However, no studies have reported on the detection of EBV DNA in saliva of NPC patients. Moreover, different DNA extraction techniques for PCR in saliva samples have never been studied. This study aimed to compare the effectiveness of DNA extraction using a commercial extraction kit against the standard boiling technique for quantitative real time PCR (RT-PCR) assays in saliva of NPC patients. Stimulated whole saliva samples from 207 newly diagnosed NPC patients were screened by qualitative PCR for the presence of EBV DNA. Twenty-three positive samples then underwent DNA extraction followed by RT-PCR testing using primer/probe set for BALF5. Duplicate measurements were performed and reproducibility was assessed. The two DNA extraction methods were evaluated by Bland-Altman’s plot and linear regression. The relationship between salivary EBV DNA level and clinical variables was evaluated by t-test. The results of RT-PCR were reproducible for 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 × 106 copies/μl) was significantly higher than the extraction kit group (1.15 ± 2.66 × 106 copies/μl) (p < 0.05). The EBV DNA level was marginally higher in patients of advanced overall stage (p = 0.05). This study showed that the performance of extraction kit was inferior to the simple boiling technique for the detection of salivary EBV DNA in NPC patients using real-time PCR. Further studies are required to enhance the detection rate of EBV DNA in saliva of NPC patients.
Persistent Identifierhttp://hdl.handle.net/10722/94380
ISSN
2015 Impact Factor: 4.286
2015 SCImago Journal Rankings: 1.764

 

DC FieldValueLanguage
dc.contributor.authorPow, EHNen_HK
dc.contributor.authorShan, JJWen_HK
dc.contributor.authorTsang, PCSen_HK
dc.contributor.authorMcMillan, ASen_HK
dc.contributor.authorKwong, DLWen_HK
dc.date.accessioned2010-09-25T15:29:41Z-
dc.date.available2010-09-25T15:29:41Z-
dc.date.issued2009en_HK
dc.identifier.citationThe 2nd World Congress of the International Academy of Oral Oncology "Oral Cancer: Imagine the Future", Toronto, Canada, 8-11 July 2009. In Oral Oncology Supplement, 2009, v. 3 n. 1, p. 200 Abstract no. P2.120en_HK
dc.identifier.issn1368-8375en_HK
dc.identifier.urihttp://hdl.handle.net/10722/94380-
dc.description.abstractNasopharyngeal carcinoma (NPC) is common in southern Chinese. It is closely associated with Epstein-Barr virus (EBV). Recent studies suggest that tumor burden is proportional to the level of EBV DNA in blood of NPC patients and that rapid blood testing can be used to guide therapeutic intervention. Instead of collecting invasive blood samples, saliva may be a viable alternative. However, no studies have reported on the detection of EBV DNA in saliva of NPC patients. Moreover, different DNA extraction techniques for PCR in saliva samples have never been studied. This study aimed to compare the effectiveness of DNA extraction using a commercial extraction kit against the standard boiling technique for quantitative real time PCR (RT-PCR) assays in saliva of NPC patients. Stimulated whole saliva samples from 207 newly diagnosed NPC patients were screened by qualitative PCR for the presence of EBV DNA. Twenty-three positive samples then underwent DNA extraction followed by RT-PCR testing using primer/probe set for BALF5. Duplicate measurements were performed and reproducibility was assessed. The two DNA extraction methods were evaluated by Bland-Altman’s plot and linear regression. The relationship between salivary EBV DNA level and clinical variables was evaluated by t-test. The results of RT-PCR were reproducible for 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 × 106 copies/μl) was significantly higher than the extraction kit group (1.15 ± 2.66 × 106 copies/μl) (p < 0.05). The EBV DNA level was marginally higher in patients of advanced overall stage (p = 0.05). This study showed that the performance of extraction kit was inferior to the simple boiling technique for the detection of salivary EBV DNA in NPC patients using real-time PCR. Further studies are required to enhance the detection rate of EBV DNA in saliva of NPC patients.-
dc.languageengen_HK
dc.publisherPergamon. The Journal's web site is located at http://www.elsevier.com/locate/oraloncologyen_HK
dc.relation.ispartofOral Oncology Supplementen_HK
dc.titleA comparison of laboratory extraction techniques for the detection of Epstein-Barr virus in saliva of nasopharyngeal carcinoma patientsen_HK
dc.typeConference_Paperen_HK
dc.identifier.openurlhttp://library.hku.hk:4550/resserv?sid=HKU:IR&issn=1368-8375&volume=3&spage=200&epage=&date=2009&atitle=A+comparison+of+laboratory+extraction+techniques+for+the+detection+of+Epstein-Barr+virus+in+saliva+of+nasopharyngeal+carcinoma+patients.en_HK
dc.identifier.emailPow, EHN: ehnpow@HKUCC.hku.hken_HK
dc.identifier.emailTsang, PCS: csptsang@hkucc.hku.hken_HK
dc.identifier.emailMcMillan, AS: annemcmillan@hku.hken_HK
dc.identifier.emailKwong, DLW: dlwkwong@hkucc.hku.hken_HK
dc.identifier.authorityPow, EHN=rp00030en_HK
dc.identifier.authorityTsang, PCS=rp00026en_HK
dc.identifier.authorityMcMillan, AS=rp00014en_HK
dc.identifier.authorityKwong, DLW=rp00414en_HK
dc.description.naturelink_to_subscribed_fulltext-
dc.identifier.doi10.1016/j.oos.2009.06.523-
dc.identifier.hkuros158657en_HK
dc.identifier.volume3en_HK
dc.identifier.spage200en_HK

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