Detection of HBV Resistance to Lamivudine in Patients with Chronic Hepatitis B Using Zip Nucleic Acid Probes in Kerman, Southeast of Iran

  • Published : 2012.08.31


HBV infection is contagious and may be transmitted vertically or horizontally by blood products and body secretions. Over 50% of Iranian carriers have contracted the infection prenatally, making this the most likely route of transmission of HBV in Iran. This study assesses the resistance to Lamivudine in patients with chronic hepatitis B infection using a new ZNA probe Real Time PCR method. To evaluate the effectiveness of Lamivudine therapy for chronic hepatitis B infection, a study was conducted on 70 patients (63 men and 7women), who had received the drug first line. All patients were tested for the presence of HBsAg and HBeAg, the serum ALT level and the HBV DNA load before and after treatment. In all samples resistance to Lamivudine was tested with the ZNA Probe. Our results showed that ZNA Probe Real Time PCR method could detect wild type,YMDD, and its mutants, tyrosine-isoleucine-aspartate-aspartate and tyrosine-valine-aspartate-Aspartate. Among an estimated seventy patients with chronic hepatitis B infection, 18 (25.7%) were resistant to lamivudine. Only one patient was negative for presence of HBS-Ag (5.6%) and two patients were negative for HBe-Ag (11.1%). Real-time PCR with Zip nucleic acid probes is a sensitive, specific and rapid detection method for mutations in the YMDD motif, which will be essential for monitoring patients undergoing Lamivudine antiviral therapy.


  1. Aberle SW, Kletzmayr J, Watschinger B, et al (2001). Comparison of sequence analysis and the inno-lipa hbv dr line probe assay for detection of lamivudine-resistant hepatitis b virus strains in patients under various clinical conditions. J Clin Microbiol, 39, 1972-4.
  2. Aghasadeghi MR, Bahramali G, Sadat SM, et al (2011). Detection of hepatitis b virus variants in hbv monoinfected and hbv/hiv coinfected iranian patients under lamivudine treatment. Curr HIV Res, 9, 263-9.
  3. Amini-Bavil-Olyaee S, Hosseini SY, Sabahi F, Alavian SM (2008). Hepatitis B virus genotype and ymdd motif mutation profile among patients infected with hbv and untreated with lamivudine. Int J Infectious Diseases, 12, 83-7.
  4. Arslan U, Ural O, Findik D (2008). YMDD motif variants detected by inno-lipa hbv assay in chronic hepatitis b patients during lamivudine therapy. Mikrobiyol Bul, 42, 445-50.
  5. Brunelle MN, Jacquard AC, Pichoud C, et al (2005). Susceptibility to antivirals of a human hbv strain with mutations conferring resistance to both lamivudine and adefovir. Hepatology, 41, 1391-8.
  6. Buti M, Elefsiniotis I, Esteban R (2007). Adefovir for lamivudine resistant hbv: More than meets the eye - reply. J Hepatology, 47, 619-20.
  7. Chen CH, Lee CM, Tung WC, et al (2010). Evolution of fulllength hbv sequences in chronic hepatitis b patients with sequential lamivudine and adefovir dipivoxil resistance. J Hepatology, 52, 478-85.
  8. Degertekin B, Hussain M, Tan J, Oberhelman K, Lok AS (2009). Sensitivity and accuracy of an updated line probe assay (hbv dr v.3) in detecting mutations associated with hepatitis b antiviral resistance. J Hepatology, 50, 42-8.
  9. Elefsiniotis IS, Paparizos V, Dourakis SP, et al (2005). Clinical outcome of long-term lamivudine treated chronic hbv infected and hiv/hbv coinfected patients. J Clinical Virology, 33, 80-2.
  10. Fallahian F, Alavian S, Kayvani H, Alaeddini F, Zamani F (2010). Lamivudine resistance in iranian chronic hepatitis b patients. Shiraz E-Medical J, 11, 63-73.
  11. Fasano M, Lampertico P, Marzano A, et al (2012). HBV DNA suppression and hbsag clearance in hbeag negative chronic hepatitis b patients on lamivudine therapy for over years. J Hepatol, 56, 1254-8.
  12. Geng H, Hua B, Wang H, et al (2006). Dual-probe assay for detection of lamivudine-resistance hepatitis b virus by realtime pcr. J Virological Methods, 132, 25-31.
  13. Kamar N, Sandres-Saune K, Ribes D, et al (2004). Effects of long-term lamivudine therapy in renal-transplant patients. J Clinical Virology, 31, 298-303.
  14. Li ZG, Chen LY, Huang J, et al (2005). Quantification of the relative levels of wild-type and lamivudine-resistant mutant virus in serum of hbv-infected patients using microarray. J Viral Hepat, 12, 168-75.
  15. Liu H, Mao R, Fan L, et al ( 2011). Detection of lamivudine- or adefovir-resistant hepatitis b virus mutations by a liquid array. J Virological Methods, 175, 1-6.
  16. Liu YC, Zhang WL, Hu Y, et al (2010). Detection of hbv resistant mutations related to lamivudine, adefovir and entecavir by reverse hybridization technique. Cma J, 18, 414-8.
  17. Moosavy SH, Froutan H, Andrabi Y, et al (2011). Frequency of ymdd mutations in patients with chronic hepatitis b untreated with antiviral medicines. Med J Islamic Republic of Iran, 25, 186-93.
  18. Noir RG, Kotera M, Pons B, Remy JS, Behr JP (2008). Oligonucleotide-oligospermine conjugates (zip nucleic acids): A convenient means of finely tuning hybridization temperatures. J Am Chem Soc, 9, 13500-5.
  19. Rizzetto M, Tassopoulos NC, Goldin RD, et al (2005). Extended lamivudine treatment in patients with hbeag-negative chronic hepatitis b. J Hepatology, 42, 173-9.
  20. Voirin E, Behr JP, Kotera M (2007). Versatile synthesis of oligodeoxyribonucleotide oligospermine conjugates. Nature Protocols, 2, 1360-8.
  21. Wu F, Wu MJ, Zhuge XL, Zhu SM, Zhu B ( 2012). Correlation of the occurrence of YMDD mutations with hbv genotypes, hbv-DNA levels, and hbeag status in chinese patients with chronic hepatitis b during lamivudine treatment. Pancreatic Diseases Int, 11, 172-6.
  22. Zhou YB, Wang YF, Zhang Y, et al (2012). In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis b virus isolates. Eur J Pharmacol, 683, 10-5.

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