Volume 11, Issue 1 (Jan-Feb- 2017 2017)                   mljgoums 2017, 11(1): 12-15 | Back to browse issues page


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Hajiahmadi N, Moradi A, Javid N, Tabarraei A. Limit of Detection of HEV RNA for Diagnosis of HEV Infection. mljgoums. 2017; 11 (1) :12-15
URL: http://mlj.goums.ac.ir/article-1-925-en.html
1- Infectious Diseases Research Center
2- Infectious Diseases Research Center , tabarraei@goums.ac.ir
Abstract:   (7641 Views)
ABSTRACT
            Background and Objectives: Diagnosis of hepatitis E virus (HEV) infection could be missed in some cases if serological tests are used solely. Molecular characterization of HEV is essential for diagnosis of acute and chronic HEV infections, and evaluating the chronic HEV infection status in immunocompromised patients. The aim of this study was to prepare a suitable HEV positive control, determine the limit of detection (LOD) of HEV RNA for a specific molecular test, and evaluate the efficiency and precision of the test.
           Methods: Genomic region of HEV NCBI reference sequence was constructed. LOD, intra-assay precision, and inter-assay precision were calculated to evaluate the efficiency and precision of the test. Then, tenfold serial dilutions of the HEV positive control were prepared. Real time PCR was performed three times for each dilution. Mean, standard deviation, and coefficient of variation of cycle thresholds obtained in three independent and simultaneous tests were calculated, and the results were analyzed.
          Results: The LOD of this test was determined as 1.4×104 copy/ml or 42 copy/reaction or 14 copy/µl. Intra-assay precision and inter-assay precision for all assays were lower than 2.5% and 10%, respectively.
          Conclusion: We propose that the real time PCR assay targeting the ORF2/3 overlapping conserved region is suitable for detection of a wide range of different HEV genotypes found in acute and chronic HEV infections. However, the precision of the test should be improved for detecting HEV RNA lower than 103 copy/ml.
          Keywords: Hepatitis E virus, Limit of Detection, Real Time PCR.
Full-Text [PDF 400 kb]   (945 Downloads)    
Type of Study: Original Paper |
Received: 2017/01/28 | Accepted: 2017/01/28 | Published: 2017/01/28 | ePublished: 2017/01/28

References
1. Hosseini-Moghaddam SM, Zarei A, Alavian SM, Mansouri M. Hepatitis E virus infection: a general review with a focus on hemodialysis and kidney transplant patients. Am J Nephrol. 2010; 31(5): 398-407. doi: 10.1159/000294505. [DOI:10.1159/000294505]
2. Ahmad I, Holla RP, Jameel S. Molecular virology of hepatitis E virus. Virus Res. 2011; 161(1): 47-58. doi: 10.1016/j.virusres.2011.02.011. [DOI:10.1016/j.virusres.2011.02.011]
3. Pérez-Gracia MT, García M, Suay B, Mateos-Lindemann ML. Current Knowledge on Hepatitis E. J Clin Transl Hepatol. 2015; 3(2): 117-26.
4. Moradi A, Besharat S, Minaiifar M, Roshandel G, Tabaraii A. Seroepidemiologic assessment of Hepatitis E virus in women of reproductive age, Gorgan. Zahedan J Res Med Sci. 2010; 12(1).
5. Tabarraei A (last), Moradi A, Rodgari D, Javid N, Bakhshandeh Nosrat. Anti-Hepatitis E Virus seroprevalence in pregnant women, in Gorgan, Iran, North East of Caspian Sea.International Conference on life science and Technology. 2011; 3: 162-4.
6. Kamar N, Dalton HR, Abravanel F, Izopet J. Hepatitis E virus infection. Clin Microbiol Rev. 2014; 27(1): 116-38. [DOI:10.1128/CMR.00057-13]
7. Mokhtari C, Marchadier E, Haïm-Boukobza S, Jeblaoui A, Tessé S, Savary J, et al. Comparison of real time RT-PCR assays for hepatitis E virus RNA detection. J Clin Virol Off Publ Pan Am Soc Clin Virol. 2013; 58(1): 36-40. [DOI:10.1016/j.jcv.2013.06.038]
8. Gerber PF, Xiao C-T, Cao D, Meng X-J, Opriessnig T. Comparison of real-time reverse transcriptase PCR assays for detection of swine hepatitis E virus in fecal samples. J Clin Microbiol. 2014; 52(4): 1045-51. [DOI:10.1128/JCM.03118-13]
9. Baylis SA, Blümel J, Mizusawa S, Matsubayashi K, Sakata H, Okada Y, et al. World Health Organization International Standard to Harmonize Assays for Detection of Hepatitis E Virus RNA. Emerg Infect Dis. 2013; 19(5): 729-35. [DOI:10.3201/eid1905.121845]
10. Hassing RJ, van der Eijk AA, Lopes VB, Snijdewind IJ, de Man RA, Pas SD, et al. Hepatitis E prevalence among HIV infected patients with elevated liver enzymes in the Netherlands. J Clin Virol Off Publ Pan Am Soc Clin Virol. 2014; 60(4): 408-10. [DOI:10.1016/j.jcv.2014.05.009]
11. Feldt T, Sarfo FS, Zoufaly A, Phillips RO, Burchard G, van Lunzen J, et al. Hepatitis E virus infections in HIV-infected patients in Ghana and Cameroon. J Clin Virol Off Publ Pan Am Soc Clin Virol. 2013; 58(1): 18-23. [DOI:10.1016/j.jcv.2013.05.004]
12. Franciscus A. HCV Viral load tests. Hepat C Basics. 2006; 2: 1-3.

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