TT Virus DNA Research Articles

The Association of Prevalence and Genotypes of Torque Teno Virus (TTV) Among Chronic Hepatitis B and C Patients in Ahvaz, 2012

Background: Torque teno virus (TTV) belongs to the Circoviridae family and was separated from Japanese patients with elevated levels of serum alanine transferase for the first time. Following this, extensive studies have been carried out around the world. Objectives: The present study was designed to investigate and determine the frequency of TTV genotypes in patients with chronic hepatitis B and C in Ahvaz city in 2012. Patients and Methods: Samples from 30 patients with chronic hepatitis C and 30 patients with chronic hepatitis B were studied and compared with 30 healthy subjects as controls. TT virus DNA was investigated using semi-nested PCR. The basic primers used were 5’ UTR. Results: Out of the 90 specimens tested, nine samples (10%) were TTV-DNA positive. Of the hepatitis C samples, four samples (13.3%) were positive. Of the hepatitis B samples, two (6.67%) were positive and 3 (10%) of the control group samples were positive. Both tth4 and tth31 isolates were separated. Conclusions: TTV is a DNA virus, but its genetic diversity is increasing. The PCR methods used were not efficient enough and were not able to identify the types of TTV in blood. The frequency obtained from our study is inconsistent with some countries but is consistent with a study conducted in India. In general, a relationship between sex and age and the prevalence of TTV was not found, and this result is the same as most other studies.

Molecular detection of TT virus and SEN virus infections in hemodialysed patients and blood donors in south of Iran

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. 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. 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. 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.

TT virus prevalence, viral loads and genotypic variability in saliva from healthy Japanese children

TT virus (TTV) is genetically variable and widespread without apparent pathogenicity; however, its epidemiological features in children were not fully understood, partly because blood sampling is often unacceptable for healthy children. We therefore used saliva specimens to investigate epidemiology of TTV infection in early childhood. Saliva samples were collected from 83 1-month-old, 110 4-month-old and 49 42-month-old children. Peripheral blood mononuclear cells (PBMC) and saliva samples were obtained in pairs from 19 healthy adults aged 40 +/- 7 years. TTV DNA was detected and quantified by real-time PCR and classified into five genogroups (G1-G5) by a series of PCRs using genogroup-specific primer pairs. TTV DNA was detected in 6, 34 and 90% of children aged 1, 4 and 42 months, respectively, and in 84% of adults. Comparable levels of TTV DNA were detected in pairs of saliva and PBMC. TTV loads in saliva were much higher in children than in adults. G3 was the most common genogroup in all age groups. The second most prevalent was G4 at 1-4 months of age and G1 thereafter. The prevalence of TTV infection reached a plateau at or before 42 months; however, somehow different epidemiologic features were observed among genogroups.

TTV infection in colorectal cancer tissues and normal mucosa

Department of Medicine II, Medical Faculty Mannheim, University of Heidelberg and the Molecular Gastroenterology Unit,Deutsches Krebsforschungszentrum, Heidelberg, GermanyDear Sir,Genetic modifications and environmental factors are presentlyseen as the key factors in the etiology of colorectal cancer,

TT Virus Infection: Role of Interferons, Interleukin-28 and 29, Cytokines and Antiviral Proteins

In 1997 a novel virus in the serum of a patient with acute post-transfusion hepatitis of non A-G etiology was identified. This agent was designed TT virus (TTV). It produces persistent viremia and no disease, but the mechanism of its persistence is poorly understood. In the present study mRNA expression of antiviral proteins as MxA, 2' 5' OAS, anti-apopotic protein, cytokines IL- 28, IL- 29 and IFN are examined in a subject affected by B lymphoma and positive for TTV DNA and RNA in this cellular subset, and in BJAB and Dohh2 cell lines.

O.098 Changes in NK cell repertoire after therapeutic HBV vaccination could be correlated to induction of HBV-specific T-cell responses

To assess the prevalence of TT virus (TTV) infection in a series of patients with chronic hepatitis C virus (HCV) infection, with or without benign (mixed cryoglobulinemia) or malignant (B-cell non-Hodgkin lymphoma (B-NHL)) lymphoproliferative disease.Sixty-six HCV patients were studied, including patients with mixed cryoglobulinemia (n=30), B-NHL (n=15), and no mixed cryoglobulinemia or B-NHL (n=21). All HCV patients had increased transaminase levels and were HCV RNA positive. Patients were considered to have mixed cryoglobulinemia if two successive determinations of their serum cryoglobulin level were above 0.05 g/L. Mixed cryoglobulinemia-negative patients never had mixed cryoglobulins in their serum on multiple determinations. Subjects without HCV infection included 79 patients with histologically proven B-NHL, and 50 healthy blood donors. Serum samples were analyzed for TTV DNA by nested polymerase chain reaction, with two couples of primers in different regions of the genome, in two independent laboratories.In the group of HCV-positive patients, TTV DNA was found in one of 15 (6.7%) patients with B-NHL, and in nine of 51 (17.6%, P=0.43) of those without B-NHL. Among HCV-positive patients without B-NHL, TTV DNA was more frequently found in those with type II mixed cryoglobulinemia vasculitis than in those without it (six of 16 (37.5%) versustwoof 21 (9.5%),P=0.05). In subjects without HCV infection, TTVDNA was present in 10 of 79 (12.7%) patients with B-NHL and in seven of 50 (14.0%, P=0.82) blood donors.In patients chronically infected with HCV, TTV co-infection: (1) is not associated with the presence of B-NHL; and (2) is more frequently found in patients presenting a type II mixed cryoglobulinemia vasculitis.

Association of Cytomegalovirus with Infantile Hepatitis

Infantile hepatitis is occasionally seen in apparently healthy children. In most cases, the etiology of the infection is uncertain. However, cytomegalovirus (CMV), Epstein-Barr virus (EBV), human herpesvirus-6 (HHV-6), human herpesvirus-7 (HHV-7), human parvovirus B19, and TT virus (TTV) are considered to be associated with hepatitis in children. The objective of this study was to investigate the correlations between these viruses and infantile hepatitis. Twenty-six children from 1 to 24 months old (median age, 7 months) who had liver dysfunction of unknown etiology were enrolled in this study. Plasma samples were examined by a real-time PCR assay for CMV, EBV, HHV-6, HHV-7, parvovirus B19, and TTV DNA. The DNA of CMV was detected in the plasma of four patients (15.4%) and was detected significantly more often in the patient group than in the control group. The CMV-infected patients were 1 to 3 months old, which was significantly younger than the remaining patients. The serological findings did not always correlate with the results of the real-time PCR assay. The DNA of TTV was detected in four patients (15.4%), while human parvovirus B19 DNA was detected in three (11.5%). However, the detection frequencies of these viral DNAs were not significantly different from those in the control groups, and some of these patients had co-infections. These results indicate that CMV might be one of the major pathogens responsible for infantile hepatitis; however, serological tests have limited utility for the diagnosis of CMV infection in young children.

Detection of TT Virus-DNA among Children with Collagen Vascular Diseases Receiving Blood Transfusions

Detection of TT Virus-DNA among Children with Collagen Vascular Diseases Receiving Blood Transfusions

Molecular Epidemiology and Clinical Implications of TT Virus (TTV) Infection in Indian Subjects

This study was aimed at obtaining data on the epidemiology and clinical course of TT virus (TTV) infections among Indian subjects. The TTV is a nonenveloped DNA virus, first identified in the peripheral blood of individuals with posttransfusion hepatitis of unknown etiology. There has been much conjecture regarding the disease association of this virus. A total of 494 serum specimens from various groups of high-risk and control subjects were screened for TTV DNA by a semi-nested PCR, using the ORF1-derived N22 primers. The sera were also screened for the HBsAg surface antigen by an ELISA, HCV RNA by a 5' NCR-based RT-PCR and GBV-C/HGV RNA by a 5' UTR-based RT-PCR. The clinical and hepatic profiles of the various subjects were also studied. Seventy-one randomly picked TTV isolates were directly sequenced and their phylogeny was studied. TTV showed an overall positivity rate of 45.34% with a significant higher prevalence of 52.9% among the high-risk subjects as against a prevalence of 28% among healthy control subjects (P < 0.001). Abnormal liver function profiles were frequent among TTV viremic individuals and among the acute hepatitis cases studied a higher mortality rate correlated with a superimposed TTV infection. The 71 TTV isolates sequenced were found to belong to genotype 1a being closely homologous to TTV prototype TA278. The TT virus shows a significant prevalence in the Indian population, particularly among subjects at risk for acquiring parenterally transmitted infections. Our study corroborates a putative role of the virus in the etiology of liver disease, particularly in coinfection with other agents.

Co-infections with Hepatitis G and TT Virus in Patients with Chronic Hepatitis C in Hungary

The significance of co-infections with novel hepatitis viruses Hepatitis G (GBV-C, HGV) and TT virus (TTV) in chronic hepatitis C is not clear. We determined the prevalence of HGV RNA and TTV DNA in chronic hepatitis C patients and in asymptomatic hepatitis C virus (HCV) carriers, and assessed the influence of these agents on the course of HCV infection. Seventy-seven patients with chronic hepatitis C--50 of them treated with interferon (IFN)--and 33 HCV carriers with normal alanine aminotransferase have been investigated. Previous HBV infection was detected by testing serum HBsAg and aHBc. HGV RNA and TTV DNA were detected by PCR. In the healthy population, the prevalence of anti-HCV was 0.3%, HGV RNA 8.0% and TTV DNA 18.5%. In chronic hepatitis C HGV RNA occurred in 9.09% and TTV DNA in 40.25% of cases. In IFN-treated patients with sustained remission, the frequency of TTV was 20% vs. 45.7% found in non-responders. Among asymptomatic HCV-carriers, the prevalence of HGV RNA was 9.09% and TTV DNA 75.7%. Neither HGV RNA nor TTV DNA had apparent effect on the HCV infection. TTV was detected with the lowest frequency in persons with sustained remission due to IFN, suggesting antiviral effect of IFN on TTV.

Molecular characterization of porcine TT virus, an orphan virus, in pigs from six different countries

Human TT virus (TTV), originally isolated from a patient with post-transfusion hepatitis in 1997, is ubiquitous and non-pathogenic. Viruses related to human TTV have since been identified in non-human primates, bovine, ovine, porcine, feline, and canine. The objective of this study was to genetically characterize porcine TTV from pigs in different geographic regions. PCR primers based on the non-coding region of the only available porcine TTV isolate were designed to amplify porcine TTV DNA from sera of pigs in six different countries. Porcine TTV DNA was detected in 66.2% (102/154) of the swine sera. The percentages of positive pigs varied greatly from country to country and even within the same country: 33% in Iowa, USA; 40% in Thailand; 46% in Ontario, Canada; 80% in China; 85% in Korea; 90% in Spain; 100% in Quebec and Saskatchewan, Canada. A total of 40 porcine TTV isolates (five from each geographic region) were sequenced for a 218 bp fragment within the non-coding region. Sequence analyses revealed that porcine TTV isolates from different geographic regions shared 86–100% nucleotide sequence identity to each other. The prototype Japanese isolate of porcine TTV, Sd-TTV31, shared 90–97% nucleotide sequence identity with porcine TTV isolates reported in this study. Phylogenetic analysis showed that the clustering of the porcine TTV isolates is not associated with geographic origins. Although porcine TTV is not known to be associated with any swine disease, co-infection of pigs with TTV and other known swine pathogens may result in enhanced disease. There are also concerns for risk of potential human infection during xenotransplantation.

TT Virus Infection of Periodontal Tissues: A Controlled Clinical and Laboratory Pilot Study

A novel single-strand, circular DNA virus has been recently isolated and named TT virus (TTV). It has been demonstrated that peripheral blood cells harbor TTV DNA, suggesting that the virus might replicate in lymphoid cells and contribute to lymphocyte imbalances with consequent immunosuppressive effects. The purpose of this study was to investigate the prevalence of TTV DNA in healthy and periodontally compromised subjects, evaluating the presence of the virus in the gingiva and saliva, and comparing virological results with clinical data. Twenty-one patients (seven males and 14 females, aged 25 to 76 years) were enrolled in the study. Eleven subjects were diagnosed with moderate periodontitis, while 10 were periodontally healthy. A sample of saliva was taken from each patient before recording the periodontal data; subsequently, a gingival biopsy was performed. A real-time polymerase chain reaction was used to quantify the presence of TTV DNA in saliva and gingival specimens. A statistically significant association was found between TTV in gingival tissue and the presence of periodontitis (P = 0.0351), while no association was observed between TTV in saliva and the presence of periodontitis (P = 0.4762). A new DNA virus (TTV) was first identified in the gingival tissue and was found to be significantly associated with the presence of periodontitis. These findings need to be investigated in further studies.

Helicobacter pylori and TT virus prevalence in Japanese children.

The major transmission route of Helicobacter pylori, oral-oral or fecal-oral, remains to be established. TT virus (TTV), a recently discovered microbe that is prevalent in healthy persons, is believed to be mainly transmitted by nonparenteral routes. The purpose of this study was to test the hypothesis that these two microorganisms have a common mode of transmission. We investigated the seroprevalence of H. pylori and TTV in a cross-sectional study of 454 healthy Japanese children from birth to age 15 years, living in five different geographic areas. Determination of H. pylori status was based on the presence of specific serum IgG and IgA antibodies, determined using enzyme immunoassays. TTV DNA was detected and the titer was determined using semiquantitative polymerase chain reaction with heminested primers. The overall prevalences of H. pylori and TTV were 12.2% and 21.6%, respectively. An age-related increase of prevalence was shown for H. pylori ( P < 0.001), but not for TTV ( P = 0.23). Titers of TTV DNA significantly decreased with age (P = 0.02). There were significant geographic differences in TTV prevalence ( P < 0.001), but not in H. pylori seroprevalence (P = 0.33). There was no true correlation between the prevalence of these two organisms (Phi coefficient = -0.02 and P = 0.66). Although Japanese children frequently acquire both H. pylori and TTV, especially in early childhood, their acquisition appears to be independent.

Distribution of TT virus genomic groups 1–5 in Brazilian blood donors, HBV carriers, and HIV‐1‐infected patients

Human isolates of the highly prevalent TT virus (TTV) have been classified into five major genomic groups (1-5). The geographical distribution of the groups throughout the world is not well known. Five different PCR assays were developed in an attempt to amplify specifically TTV DNAs of each genomic group. Serum samples collected from 72 Brazilian adults (24 voluntary blood donors, 24 hepatitis B virus (HBV) carriers, and 24 human immunodeficiency virus type 1 (HIV-1)-infected patients) were tested. TTV DNA from at least one genomic group was detected in 11 (46%) blood donors, 13 (54%) HBV carriers, and 24 (100%) HIV-1 patients. All five genomic groups were detected in the three populations, with the exception of group 2 in blood donors. Some samples, negative with all five specific assays, were positive with the commonly used untranslated region (UTR) PCR system. On the other hand, TTV DNA was detected in some samples by using specific assays but not with the UTR PCR. Mixed infections with 2-5 TTV isolates from different groups were detected in 21% blood donors, 29% HBV carriers, and 71% HIV-1 patients. Fifteen PCR products (three obtained with each assay) were sequenced. Most sequences showed high (>86%) homology with those of TTV isolates belonging to their presumed groups. However, three sequences had low homology with all TTV sequences available from the DNA databanks. In conclusion, TTV isolates belonging to all five known genomic groups circulate in Brazil, and the results suggest the existence of new and as yet uncharacterised major genomic groups.

Clinical significance of TT virus (TTV) infection in chronic hepatitis C patients with high dose interferon-alpha therapy in Taiwan: re-evaluated by using new set of TTV primers

Background: The clinical significance of TT virus (TTV) coinfection in chronic hepatitis C (CHC) patients and influence of TTV viremia on hepatitis C virus (HCV) response to high dose interferon-alpha therapy in Taiwan were investigated. Materials and Methods: Total 102 HCV RNA-positive CHC patients were enrolled. TTV DNA (using polymerase chain reaction primers derived from 5′ non-coding region and open reading frame 2), alanine aminotransferase (ALT), GB virus-C/hepatitis G virus (GBV-C/HGV) RNA, anti-E2 antibody, genotype and RNA levels of HCV were tested. Results: The prevalence of TTV DNA was 51.0%. The mean age of TTV viremic CHC patients was significant higher than non-viremic ones ( P<0.05). HCV sustained viral response (SVR) was achieved in 42 (41.2%) patients. Based on multivariate regression analyses, SVR were significantly associated with low pretreatment HCV RNA levels, HCV genotype non-1b and high pretreatment levels of ALT but not TTV viremia. Conclusions: TTV viremia is highly prevalent among Taiwanese CHC patients and related to increased ages. Neither severity of liver disease nor replication and genotype distribution of HCV was affected by concurrent TTV infection. With high HCV SVR rate associated with pretreatment HCV RNA and ALT levels and HCV genotype, TTV viremia did not influence the HCV response.

Detection of TT virus in lymph node biopsies of B-cell lymphoma and Hodgkin's disease, and its association with EBV infection.

Human TT virus (TTV) recently isolated from the serum of a patient with post-transfusion hepatitis does seem to have only hepatopathic effect. The virus can also infect the serum, peripheral blood mononuclear cells (PBMC) and bone marrow cells (BMC ). Additional evidence has indicated that TTV is also present in the serum of people with hematopoietic malignancies. A significant increase in the incidence of lymphoma has recently been observed worldwide. We have investigated the presence of TTV DNA in lymph node biopsies of Italian patients affected with the most common lymphoma types in Western Countries: follicular lymphoma (FL), diffuse large B-cell lymphoma (DLBCL) and nodular sclerosis Hodgkin's disease (NS-HD). The possible role of a co-infection with Epstein-Barr virus (EBV) has also been investigated. DNA was extracted from 73 paraffin-embedded and 38 snap-frozen tissue specimens. From these, only 67 samples (29 paraffin-embedded and 38 snap-frozen tissues) from a total of 56 patients, were suitable for PCR analysis. TTV and EBV were detected by PCR using primers from two different conserved region in TTV and EBV genomes respectively. TTV DNA was detected in 30.0-50.0% of FL, 30.8% of DLBCL and 30.0-50.0% of NS-HD cases, depending on the primers used. All cases of non-specific reactive lymphoid hyperplasia (RLH), used as a putative control, were negative. The two major TTV genotypes circulating in Italy (G1 and G2) were detected in the analysed lymphoid neoplasms. EBV DNA was detected in 40.0% of FL, in 72.7%of DLBCL, in 80.0% of SN-HD and in 40.0% of RLH cases. EBV co-infection was found in 90% of TTV positive cases. The in situ hybridization assay was performed in TTV positive frozen samples. The significant prevalence of TTV DNA in lymphocytes circulating in the lymph nodes of both B-cell lymphomas and HD reported herewith suggests an implication of TTV infection in the development of these lymphoproliferative disorders.

TT Virus Infection Does Not Affect the Clinical Profiles of Patients with Hepatitis B and C in Yanbian City, China

China is an area of high endemicity for viral hepatitis, and the molecular epidemiological investigation of TT virus (TTV) infection is of interest. In the present study, we investigated the epidemiology, clinical significance and molecular characteristics of TTV infection in patients with chronic hepatitis B and C in Yanbian City, China. Serum samples obtained from 74 patients with hepatitis B and hepatitis C who visited Yanbian Hospital, located in northeast China, were analyzed in this study. The study group included 22 cases of chronic hepatitis B (B-CH), 17 cases of liver cirrhosis B (B-LC), 7 cases of hepatocellular carcinoma (B-HCC), 16 cases of chronic hepatitis C (C-CH), 11 cases of liver cirrhosis C (C-LC) and 1 case of hepatocellular carcinoma (C-HCC). Detection of TTV DNA was performed as described by Nishizawa et al. The second-round PCR products from 7 subjects were sequenced, followed by investigation of nucleotide homology and phylogenetic analysis. TTV DNA was present in 18.2, 5.9, 28.6, 6.3, 9.1 and 0% of the patients with B-CH, B-LC, B-HCC, C-CH, C-LC and C-HCC, respectively. The highest prevalence of TTV infection was seen in the groups aged 40–50 and over 60 years. There was no significant correlation between the presence of TTV DNA and the clinical parameters in patients with hepatitis B and C. The various isolates showed 97.9–100% with isolates reported previously from Japan and 98.4–100% with isolates reported previously from China. Nucleotide sequence analysis revealed that the Yanbian isolates could be classified in the same group as the Japan and China isolates. We concluded that chronic coinfection with TTV did not affect the serological features of chronic hepatitis B and C in China, as found in Tokyo, Japan.

TT virus infection in patients with chronic hepatitis B and response of TTV to lamivudine.

To investigate the responses of TT virus (TTV) and hepatitis B virus (HBV) to a long-term lamivudine therapy. Sixteen patients infected with both TTV and HBV were treated with lamivudine 100 mg daily for 30 months. Blood samples were drawn at the beginning of the therapy and subsequently at month 3, 6, 9, 12 and 30. Serum TTV was quantified by real time PCR and serum HBV was detected by hybridization assay and nested polymerase chain reaction. TTV infection was detected in 100 % of HBV-infected patients. Loss of serum TTV DNA after one year of treatment occurred in 1/16 (6 %) patients. At the end of therapy, TTV DNA was positive in 94 % of them. The decline of HBV viremia was evident at 3 months after therapy and the response rate was 31 %, 44 %, 63 %, 50 % and 50 % at month 3, 6, 9, 12 and 30, respectively. TTV replication is not sensitive to lamivudine and is highly prevalent in HBV-infected patients.

GB virus C/hepatitis G virus and TT virus infections among high risk renal transplant recipients in India.

GB virus C/hepatitis G virus (GBV-C/HGV) and TT virus (TTV) have been widely reported in patients with high parenteral risk such as haemodialysis and renal transplant recipients. The occurrence of these agents in association with hepatitis B virus (HBV) and hepatitis C virus (HCV), in Indian renal transplant recipients, is yet unreported. Molecular and serological markers of GBV-C/HGV and TTV were examined in addition to those for HBV, HCV and hepatitis D virus (HDV) in a selected group of seventy renal transplant recipients. HGV RNA detection was achieved using primers specific for the 5'NCR and NS5a regions of the genome. Anti-GBV-C/HGV antibody was detected using the mu plate anti-HG env kit (Roche, Germany). TTV DNA PCR was performed using primers specific for the coding region (method A) of the genome. In 50% of patients, TTV DNA was also tested for using primers specific for the non-coding region (method B). Host related factors such as age, alanine aminotransferase (ALT) levels, number of transfusions, haemodialysis sessions, and months following transplantation were also studied. Exposure rates to GBV-C/HGV, TTV (method A), HBV, HCV and HDV were 58.6, 32.9, 52.9, 54.3 and 2.9%, respectively. 'Active' infection as measured by viraemia and/or virus-specific antigenaemia for GBV-C/HGV, TTV, HBV and HCV was 52.9, 32.9, 15.7 and 52.9%, respectively. The majority of GBV-C/HGV and TTV infections were seen as co-infections with other hepatitis viruses. Single infection with GBV-C/HGV and TTV was seen in ten (14.2%) and eight (11.4%) patients, and was not associated with ALT elevation when compared to uninfected blood donors. Using univariate analysis, GBV-C/HGV RNA was significantly associated with > or =20 haemodialysis sessions. TTV DNA occurrence was not associated with any risk factors. There is a high occurrence of GBV-C/HGV and TTV in this select group of renal transplant recipients in India. These viruses mostly occurred in the context of co-infections with other hepatitis viruses. Long term effects of multiple hepatotropic viral infections need to be carefully documented in such transplant populations.

Comparison of DNA and RNA extraction methods for mummified tissues

Nucleic acids extracted from mummified tissues are valuable materials for the study of ancient human beings. Significant difficulty in extracting nucleic acids from mummified tissues has been reported due to chemical modification and degradation. The goal of this study was to determine a method that is more efficient for DNA and RNA extraction from mummified tissues. Twelve mummy specimens were analyzed with 9 different nucleic acid extraction methods, including guanidium thiocyanate (GTC) and proteinase K/detergent based methods prepared in our laboratory or purchased. Glyceraldehyde 3-phosphate dehydrogenase DNA and β-actin RNA were used as markers for the presence of adequate DNA and RNA, respectively, for PCR and RT-PCR amplification. Our results show that 5 m GTC is more efficient of releasing nucleic acids from mummified tissue than proteinase K/detergent, and phenol/chloroform extraction with an additional chloroform step is more efficient than phenol/chloroform along. We were able to isolate DNAs from all 12 specimens and RNAs from 8 of 12 specimens, and the nucleic acids were sufficient for PCR and RT-PCR analysis. We further tested hepatitis viruses including hepatitis B virus, hepatitis C virus, hepatitis G virus, and TT virus DNA, and fail to detect these viruses in all 12 specimens.