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  Table of Contents    
ORIGINAL ARTICLE  
Year : 2012  |  Volume : 55  |  Issue : 4  |  Page : 478-480
Molecular detection of TT virus and SEN virus infections in hemodialysed patients and blood donors in south of Iran


1 Department of microbiology, Jahrom Branch, Young Researchers Club, Islamic Azad University, Jahrom, Iran
2 Department of microbiology, Gerash Research Center, Shiraz University of Medical Sciences, Shiraz, Iran

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Date of Web Publication4-Mar-2013
 

   Abstract 

Background: SEN virus (SEN-V) and TT virus (TTV) have been classified in the circoviridae family. Both are single-stranded, non-enveloped DNA viruses of about 3800 nucleotides. Patients on maintenance hemodialysis (HD) have a high risk of blood-borne viral infections. SEN-V and TTV has been reported from a number of HD units from various countries throughout the world. Materials and Methods: A total of 377 blood samples obtained from 150 healthy donors and 227 HD patients were collected at the HD center. SEN-V and TTV DNA was determined by polymerase chain reaction (PCR) in all samples. Results: TTV was detected in 109 (48.01%) of 227 hemodialysed patients and 14 (9.33%) of 150 voluntary blood donors (significant, P < 0.05). The PCR results for SEN-V-D/H DNA showed that 65 (28.63%) were positive for SEN-V-D and 33 (14.53%) were positive for SEN-V-H. 9.69% of 227 patients were positive for SEN-V-D/H co-infection. In the control group, SEN-V-D was detected in 14 (9.33%) and SEN-V-H was detected in 15 (10%) of the 150 (100%) blood donors. Conclusion: These findings show that the prevalence of SEN-V-D/H and TTV is higher than healthy blood donors. Also, these results indicate that the prevalence of SEN-V and TTV infections in our region is similar with that in other countries.

Keywords: Blood donors, hemodialysis, SEN-V infection, TT virus

How to cite this article:
Afkari R, Pirouzi A, Mohsenzadeh M, Azadi M, Jafari M. Molecular detection of TT virus and SEN virus infections in hemodialysed patients and blood donors in south of Iran. Indian J Pathol Microbiol 2012;55:478-80

How to cite this URL:
Afkari R, Pirouzi A, Mohsenzadeh M, Azadi M, Jafari M. Molecular detection of TT virus and SEN virus infections in hemodialysed patients and blood donors in south of Iran. Indian J Pathol Microbiol [serial online] 2012 [cited 2019 Sep 18];55:478-80. Available from: http://www.ijpmonline.org/text.asp?2012/55/4/478/107784



   Introduction Top


SEN virus (SEN-V) and TT virus (TTV) have been classified in the circoviridae family. Five hepatitis viruses (A-E) have been established for viral hepatitis cases and there are still patients with acute or chronic hepatitis with unknown origin (non-A to -E hepatitis). TTV and SEN-V are assumed to be responsible for post-transfusion non A-E hepatitis. [1],[2] TTV is a small single-stranded DNA that was first discovered in the sera of three Japanese patients with post-transfusion hepatitis in 1997. [3] Several studies have shown that TTV is distributed widely throughout the word. [4],[5],[6] SEN-V is a single-stranded DNA virus and distantly related to the TTV. Among the nine genotypes, SEN-V-D and SEN-V-H are more prevalent in serum samples from patients with transfusion-associated non A-E hepatitis. [7],[8] Parenteral transmission through blood and blood products is clearly evidenced by the higher detection rates of these viruses among multiple transfused individuals. Hemodialysed patients are at high risk for blood-borne infections because of getting regular blood for a long time and the potential for exposure to contaminated equipment. [8],[9],[10] Information regarding TTV and SEN-V infections in the Iran population is less. The aim of this study was to determine the prevalence of TTV and SEN-V in Iranian blood donors and patients on maintenance hemodialysis (HD) by molecular methods.


   Materials and Methods Top


Serum samples were collected from patients and voluntary blood donors. This study included 377 blood samples that were obtained from 150 healthy donors and 227 HD patients. These cases were recruited from HD center in Larestan Hospital, Fars, June 2007 and May 2010. Informed consent was obtained from all the patients at the initial presentation. Blood samples were centrifuged and stored at −70°C within hours of collection. Total nucleic acids from 100 μl of serum were isolated with the QIAamp blood kit, and the nucleic acid was re-suspended in 50 μl of distilled water. Polymerase chain reaction (PCR) assays were performed using 3 μl of the DNA sample, 1 × PCR buffer [10 mM Tris-HCl (pH 9.0) 50 mM KCl, 1 mM MgCl 2 , 0.01% gelatin, and 0.1% Triton X-100], 200 μM concentration of each dNTP, and 100 ng of each primer in a final volume of 25 μl.

Total DNA was amplified by semi-nested PCR using primers NG059, NG061, and NG063. The first round PCR program consisted of 35 cycles of denaturation for 30 s at 94°C, annealing for 45 s at 60°C, and extension for 45 s at 72°C, with the sense primers NG059 (5′-ACAGACAGAGGAGAAGGCAACATG-3′) and anti-sense primer NG063 (5′-CTGGCATTTTACCATTTCCAAAGTT-3′). The second round of PCR was performed with the sense primer NG061 (5′-GGCAACATGTTATGGATAGACTGG-3′) and the anti-sense primer NG063 for 25 cycles, under the same conditions as used for the first round of PCR. The amplified DNA products were analyzed by 3% agarose gel electrophoresis, stained with ethidium bromide, and photographed under UV light.

SEN-V-D/H DNA was determined by PCR. Nucleic acids were isolated from 100 μl of serum with QIAamp blood kit and re-suspended in 50 μl of elution buffer. DNA was amplified by nested PCR, with forward primer AI-1F (5′-TWCYCMAAC GAC CAG CTA GAC CT-3′; W = A or T, Y = C or T, M = A or C) and reverse primer AI-1R (5′-GTT TGT GGT GAG CAG AAC GGA-3′) for the first round for all the SEN-V genotypes. The PCR conditions in the first round were: denaturation for 45 s at 94°C, primer annealing for 45 s at 56°C, and extension for 45 s at 72°C, with a final extension step for 7 min at 72°C for 35 cycles. Second round PCR amplification was done with specific primers. Forward and reverse primers for SEN-V-D included D-1148F (5′-CTA AGC AGC CCT AAC ACTCATCCAG-3′) and D-1341R (5′-GCAGTTGACCGCAAAGTTACAAGA G-3′), and for SEN-V-H they included H-1020F (5′-TTT GGC TGC ACC TTC TGG TT-3′) and H-1138R (5′-AGA AAT GAT GGGTGAGTGTTAGGG-3′). The amplification products were separated by 3% agarose gel electrophoresis, stained with ethidium bromide, and photographed under UV light.

Statistical Analysis

Statistical analyses were conducted by SPSS software version 15.0. The differences between prevalence of SEN-V infection in hemodialysed patients and healthy blood donors were examined by t-test, Chi-square, and one-way analysis of variance (ANOVA) statistical analysis. P values < 0.05 were considered significant.


   Results Top


The details of the TTV and SEN-V-D/H in HD patients and healthy blood donors are summarized in [Table 1], [Table 2], [Table 3] and [Table 4]. Viral DNAs were extracted from 200 μl serum samples collected from hemodialysed patients and blood donors. Of the 227 hemodialysed patients, TTV was detected in 109 (48.01%) HD patients and 14 (9.33%) subjects of 150 voluntary blood donors (significant, P < 0.05). The PCR results for SEN-V-D/H DNA showed that 65 (28.63%) were positive for SEN-V-D and 33 (14.53%) were positive for SEN-V-H. 9.69% of 227 patients were positive for SEN-V-D/H co-infection. In the control group, SEN-V-D was detected in 14 (9.33%) and SEN-V-H was detected in 15 (10%) of the 150 (100%) blood donors. Among TTV-positive patients, SEN-V-D and SEN-V-H co-infections were present in 48.01% and 9.69% cases, respectively. The mean age was 44.0 ± 6.7 years for TTV-positive HD patients and 35.63 ± 1.9 years for TTV-negative HD patients (not significant, P > 0.05). The mean duration on dialysis was 14.48 ± 5.27 months for TTV-positive patients and 6.05 ± 0.97months for TTV-negative patients (significant, P < 0.05).
Table 1: Prevalence of SEN Virus (SEN - V) in Patients on maintenance haemodialysis (N=227)

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Table 2: Clinical details of the TTV-positive and TTV-negative cases (N=227)

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Table 3: Prevalence of SEN Virus (SEN - V) in voluntary blood donors (N=150)

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Table 4: Clinical details of the TTV - positive and TTV - negative cases in voluntary blood donors. (N=150)

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   Discussion Top


The aim of this study was to investigate the prevalence of SEN-V-D/H and TTV in serum of hemodialysed patients and healthy donors as controls in south of Iran. Because of the therapeutic procedures that are frequently associated with bleeding and blood transfusions, HD patients are at risk of infection by blood-borne viruses. [11] In this study, the prevalence of SEN-V-D/H DNA was 43.16% (SEN-V-D 28.63% and SEN-V-H 14.53%) and that of TTV DNA was 48.01% among hemodialysed patients. SEN-V-D/H DNA and TTV DNA were detected in 19.33% (SEN-V-D 9.33% and SEN-V-H 10%) and 9.33% of healthy blood donors, respectively. SEN-V-D/H and TTV infections were found frequently in dialysed patients rather than in healthy blood donors. It seems that the high prevalence of these groups of viruses in HD patients is due to contaminated equipments and blood products. Also, the length of HD has some effects on SEN-V and TTV positivity. The results showed that the prevalence of SEN-V and TTV in HD patients is rather high than in healthy blood donors, which was close to that reported in other studies to a high degree. In other reports, the prevalence of TTV was reported to be between 2% and 53%, [12],[13] and SEN-V-D/H was detected in 38% in Japan, 68% in northern Taiwan, and 61.6% in southern Taiwan, among hemodialysed patients. [14],[15],[16] Thus, this higher frequency implies more attention should be paid to these viruses as other blood-borne viruses.


   Acknowledgment Top


This project was supported by Shiraz University of Medical Sciences and Gerash Research Center.

 
   References Top

1.Tanaka Y, Primi D, Wang RY, Umemura T, Yeo AE, Mizokami M. Genomic and molecular evolutionary analysis of a newly identified infectious agent (SEN virus) and its relationship to the TT virus family. J Infect Dis 2001;183:359-67.  Back to cited text no. 1
    
2.Abe K, Inami T, Asano K. TT virus infection is widespread in the general populations from different geographic regions. J Clin Microbiol 1999;37:27-35.  Back to cited text no. 2
    
3.Nishizawa T, Okamoto H, Konishi K, Yoshizawa H, Miyakawa Y, Mayumi M. A novel DNA virus (TTV) associated with elevated transaminase levels in post-transfusion hepatitis of unknown etiology. Biochem Biophys Res Commun 1997;241:92-7.  Back to cited text no. 3
    
4.Inami T, Konomi N, Arakawa Y, Abe K. High prevalence of TT virus DNA in human saliva and semen. J Clin Microbiol 2000;38:2407-8.  Back to cited text no. 4
[PUBMED]    
5.Das K, Kar P, Gupta RK, Das BC. Role of transfusion.-transmitted virus in acute viral hepatitis and fulminant hepatic failure of unknown etiology. J Gastroenterol Hepatol 2004;19:406-12.  Back to cited text no. 5
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6.Okamoto HT, Nishizawa N, Kato M, Ukita H, Ikeda H, Iizuka Y, et al. Molecular cloning and characterization of a novel DNA virus (TTV) associated with post transfusion hepatitis ofunknown etiology. Hepatol Res 1988;10:1-16.  Back to cited text no. 6
    
7.Dai CY, Chuang WL, Chang WY, Chen SC, Sung MH, Hsieh MY. SEN virus infection among patients on maintenance hemodialysis in southern Taiwan. J Infect 2005;51:110-5.  Back to cited text no. 7
    
8.Mu SJ, Du J, Zhan LS. Prevalence of a newly identified SEN virus in China. World J Gastroenterol 2004;10:2402-5.  Back to cited text no. 8
    
9.Akiba J, Umemura T, Alter HJ. SEN virus: Epidemiology and characteristics of a transfusion-transmitted virus. Transfusion 2005;45:1084-8.  Back to cited text no. 9
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10.Kobayashi N, Tanaka E, Umemura T. Clinical significance of SEN virus infection in patients on maintenance hemodialysis. Nephrol Dial Transplant 2003;18:348-52.  Back to cited text no. 10
    
11.Simmonds P. TT virus infection: A novel virus-host relationship. J Med Microbiol 2002;51:455-8.  Back to cited text no. 11
[PUBMED]    
12.Forns X, Hegerich P, Darnell A. High prevalence of TT virus (TTV) infection in patients on maintenance hemodialysis: Frequent mixed infections with different genotypes and lack of evidence of associated liver disease. J Med Virol 1999;59:313-7.  Back to cited text no. 12
    
13.Utsunomiya S, Yoshioka K, Wakita T. TT virus infection in hemodialysis patients. Am J Gastroenterol 1999;94:3567-70.  Back to cited text no. 13
    
14.Jeon M, Shin J, Suh SP. TT virus and hepatitis G virus infections in Korean blood donorsand patients with chronic liver disease. World J Gastroenterol 2003;9:741-4.  Back to cited text no. 14
    
15.Kao JH, Chen W, Chen PJ, Lai MY, Chen DS. Prevalence and implication of a newly identified infectious agent (SEN virus) in Taiwan. J Infect Dis 2002;185:389-92.  Back to cited text no. 15
[PUBMED]    
16.Erensoy S, Sayiner AA, Turkoglu S, Canatan D, Akarca US, Sertöz R, et al. TT virus infection and genotype distribution in blood donors and a group of patients from Turkey. Infection 2002;30:299-302.  Back to cited text no. 16
    

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Correspondence Address:
Mohammad Jafari
Department of Shiraz University of Medical Sciences, Gerash Research Center, Gerash
Iran
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/0377-4929.107784

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    Tables

  [Table 1], [Table 2], [Table 3], [Table 4]

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