Sains Malaysiana 47(3)(2018): 531–536

http://dx.doi.org/10.17576/jsm-2018-4703-13

 

Saliva Sampling of Alcoholic Participants using Three Saliva Collection Methods

(Persampelan Air Liur menggunakan Tiga Kaedah Pengumpulan Air Liur daripada Peserta yang Mengamalkan Minuman Beralkohol)

 

KHAN, S.S., JAMEEL, R.A., RAZAK, F.A. & BAKRI, M.M.*

 

University of Malaya, 50603 Kuala Lumpur, Federal Territory, Malaysia

 

Diserahkan: 6 April 2017/Diterima: 29 September 2017

 

ABSTRACT

The potential of using saliva as a diagnostic fluid is well documented. The aim of this study was to assess the quality and quantity of saliva DNA of alcoholic and non-alcoholic participants using three saliva collection methods; DNA-SalTM (Oasis Diagnostics, USA), Oragene-DNA (DNA Genotek Inc, Ontario, Canada) and whole saliva collection method. Saliva DNA of non-alcoholic (n=30) and alcoholic participants (n=10) age between 25 and 35 years was assessed qualitatively and quantitatively using spectrophotometry. Saliva DNA quantity was the highest for all participants when using the DNA-Sal TM saliva collection kit (p<0.05). The use of a mechanical scraper provided only in the DNA-Sal TM kit may have contributed to the highest DNA yield for all participants. The quantity of saliva DNA when assessed using spectrophotometer was found to be significantly lower (p<0.05) for the alcoholic (16±3.57 ng/μL) than non-alcoholic participants (19.92±6.18 ng/μL). To determine the integrity of the DNA samples, PCR amplification of the Alcohol Dehydrogenase gene, ADH1B was carried out and the PCR was found to be successful. For all participants, the DNA quality of the saliva collected using the three saliva collection methods was found to be in the acceptable range considered as pure DNA. The DNA quality and quantity of saliva collected from the three saliva collection methods were considered suitable for research purposes.

Keywords: Alcohol; PCR; saliva; saliva collection methods

 

ABSTRAK

Potensi menggunakan air liur sebagai alat diagnostik telah pun mendapat pendedahan yang meluas. Tujuan kajian ini adalah untuk mengkaji kualiti dan kuantiti DNA menggunakan sampel air liur yang diperoleh daripada peserta yang mengamalkan minuman beralkohol. Sampel air liur diperoleh daripada semua peserta menggunakan 3 kaedah pengumpulan air liur; DNA-SalTM (Oasis Diagnostics, USA), Oragene-DNA (DNA Genotek Inc, Ontario, Canada) dan pengumpulan air liur secara langsung daripada kaviti mulut. Persampelan air liur melibatkan peserta yang tidak mengamalkan minuman beralkohol (n=30) dan yang mengamalkan minuman beralkohol (n=10) serta berumur antara 25 dan 35 tahun. Kualiti dan kuantiti DNA air liur daripada semua peserta dikaji menggunakan spektrofotometer. Kuantiti DNA air liur bagi semua peserta adalah paling tinggi apabila menggunakan kaedah pengumpulan air liur DNA-Sal TM (p<0.05). Penggunaan alat mengikis yang dibekalkan hanya untuk kaedah pengumpulan air liur DNA-Sal TM didapati berkemungkinan menyumbang terhadap kuantiti DNA yang paling tinggi. Walau bagaimanapun, pemeriksaan spektrofotometer mendapati bahawa kuantiti DNA air liur bagi peserta yang mengamalkan alkohol (16±3.57 ng/μL) adalah lebih rendah (p<0.05) berbanding dengan peserta yang tidak mengamalkan alkohol (19.92±6.18 ng/μL). Untuk memastikan integriti DNA air liur, DNA yang diperoleh daripada air liur digunakan untuk amplifikasi PCR. Amplifikasi PCR didapati telah berjaya bagi salah satu gen kumpulan Alkohol Dehidrogenase, ADH1B. Kualiti DNA air liur yang dikumpul menggunakan ketiga-tiga kaedah pengumpulan air liur bagi semua jenis peserta didapati berada dalam lingkungan yang dianggap sebagai DNA tulen. Kualiti dan kuantiti DNA bagi air liur yang dikumpul menggunakan ketiga-tiga kaedah pengumpulan air liur adalah dianggap sesuai bagi kegunaan penyelidikan.

Kata kunci: Air liur; alkohol; kaedah pengumpulan air liur; PCR

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*Pengarang untuk surat-menyurat; email: marinab@um.edu.my

 

 

 

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