Hepatit B virus DNA kantitasyonunda ABI prism 7000 ile 7700 sekans saptama cihazlarının sonuçlarının birbiri ile uygunluğunun araştırılması
Küçük Resim Yok
Tarih
2005
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Dergi ISSN
Cilt Başlığı
Yayıncı
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Tanısal viroloji laboratuvarlarında klinik anlamlılığı nedeniyle önemli olan kantitatif HBV-DNA testinde çeşitli yöntemler ve sistemler kullanılmaktadır. Bu çalışma daha önce ABI Prism 7700 cihazı (PE Biosystems) için tasarlanıp, optimize edilmiş dinamik aralığı 373-1010 kopya/ml olan polimeraz zincir reaksiyonu (PCR) protokolünün ABI 7000 (PE Biosystems) cihazındaki kullanımını değerlendirmek için planlanmıştır. Kronik B hepatitli 168 hastanın serumu çalışmaya alınmış, nükleik asit izolasyonu "High Pure Viral Nucleic Acid" kiti ile (Roche Applied Science, ABD) MagnaPure LC izolasyon istasyonunda (Roche Applied Science, Germany) yapılmıştır. ABI Prism 7000 ve 7700 cihazlarında HBV-DNA'nın pre-S gen bölgesini hedefleyen gerçek zamanlı HBV-DNA PCR protokolü uygulanmıştır. Test edilen 168 örnekten 124'ü testlerin dinamik aralık sınırları içinde saptanmıştır. Her iki cihazdan 124 örnek için elde edilen sonuçlar birbiriyle uyumlu bulunmuştur. Kalan 44 örnekten biri her iki cihazla 1010kopya/ml'nin üstünde, altı örnek sadece 7700 ile 1010kopya/ml'nin üstünde saptanmıştır. Otuz örnek her iki cihazla negatif iken, ABI 7000 ile 320-1220 kopya/ml arasında pozitif olan yedi örnek ABI Prism 7700 ile negatif sonuç vermiştir. Sonuç olarak, bu PCR protokolünün ABI 7000 cihazında referans serumlarla elde edilen çalışma sonuçlarına göre uygulanabilir olduğu, ancak HBV-DNA düzeyi 1x106 kopya/ml'nin altındaki örneklerde 7700 ile elde edilen sonuçlarla uyumsuzluk göstermesi nedeniyle aynı hastanın izleminde farklı cihazların kullanılmaması gerektiği kanısına varılmıştır.
There are different methods and systems for quantification of HBV-DNA in clinical virology laboratories. the aim of this study was to evaluate the agreement of the polymerase chain reaction (PCR) protocol with ABI Prism 7000 instrument (PE Biosystems) which was designed and optimised for ABI Prism 7700 (PE Biosystems). Serum samples obtained from 168 chronic hepatitis B patients were treated with "High Pure Viral Nucleic Acid Kit" (Roche Applied Science, USA), and MagnaPure LC isolation station (Roche Applied Science, Germany) was used for HBV-DNA isolation. Real time PCR procedure which amplifies pre-S gene of HBV genome was performed. Amplification and detection steps of all samples were performed with ABI Prism 7700 and 7000 Sequence Detection Systems. Among 168 samples, results of 124 serum samples were found to be in dynamic ranges of the tests. the results of these 124 samples obtained from ABI 7000 and ABI 7700 were concordant. Among the rest of 44 samples; one yielded higher than 1010 copies/mL with two of the systems; six samples gave results higher than 1010copies/mL only with 7700; thirty samples were found negative with both of the systems; seven samples were positive (320-1220 copies/mL) with 7000 but negative with 7700. As a result this PCR protocol can be used in ABI 7000 system according to viral quality control (VQC) results. However, since the results of samples with HBV-DNA less than 1x106 copy/ml were discordant with the results obtained by ABI 7700 system, it can be concluded that different systems must not be used for the management and monitoring of the same patient.
There are different methods and systems for quantification of HBV-DNA in clinical virology laboratories. the aim of this study was to evaluate the agreement of the polymerase chain reaction (PCR) protocol with ABI Prism 7000 instrument (PE Biosystems) which was designed and optimised for ABI Prism 7700 (PE Biosystems). Serum samples obtained from 168 chronic hepatitis B patients were treated with "High Pure Viral Nucleic Acid Kit" (Roche Applied Science, USA), and MagnaPure LC isolation station (Roche Applied Science, Germany) was used for HBV-DNA isolation. Real time PCR procedure which amplifies pre-S gene of HBV genome was performed. Amplification and detection steps of all samples were performed with ABI Prism 7700 and 7000 Sequence Detection Systems. Among 168 samples, results of 124 serum samples were found to be in dynamic ranges of the tests. the results of these 124 samples obtained from ABI 7000 and ABI 7700 were concordant. Among the rest of 44 samples; one yielded higher than 1010 copies/mL with two of the systems; six samples gave results higher than 1010copies/mL only with 7700; thirty samples were found negative with both of the systems; seven samples were positive (320-1220 copies/mL) with 7000 but negative with 7700. As a result this PCR protocol can be used in ABI 7000 system according to viral quality control (VQC) results. However, since the results of samples with HBV-DNA less than 1x106 copy/ml were discordant with the results obtained by ABI 7700 system, it can be concluded that different systems must not be used for the management and monitoring of the same patient.
Açıklama
Anahtar Kelimeler
Viroloji, Biyoteknoloji ve Uygulamalı Mikrobiyoloji, Mikrobiyoloji
Kaynak
Mikrobiyoloji Bülteni
WoS Q Değeri
Scopus Q Değeri
Cilt
39
Sayı
2
