PROSPECTIVE OBSERVATIONAL COHORT STUDY OF PREDICTORS OF THE EFFECTIVENESS OF PEGYLATED INTERFERON IN PATIENTS WITH HEPATITIS C IN GEORGIA.

The main objective of the study was to evaluate the predictive values of virological response at 4th and 12th weeks after treatment initiation on sustained virological response by HCV genotype in patients with hepatitis C in Georgia. Local, non-interventional, prospective, cohort study was conducted in 2011-2016. A cohort of adult peginterferon treatment naïve patients with chronic hepatitis C were observed during the complete active treatment period with PEGASYS®/COPEGUS® and 24 weeks after the end of treatment. HCV RNA titers were assessed prior to treatment, after 4, 12 weeks of treatment, at the end of treatment and 24 weeks after the end of treatment.530 men and women aged ≥18 years with serologically proven CHC (all genotypes) were enrolled in this study. Enrolles study subjects were treated with PEGASYS® 180mkg (peginterferon alfa-2a) in combination with COPEGUS® 200mg (ribavirin) according to the current standard of care and in line with current summary of product characteristics. All clinical information for this cohort study was collected from the patient's medical records. All laboratory parameters which were collected for this observational study according to the protocol were performed in study centers. All data were analyzed with descriptive and analytical statistics. Our analysis demonstrated that the early achievement of viral response predicts the higher probability of achieving sustainvirul response. The viral response itself was strongly associated with baseline liver fibrosis quantitive HCV RNA level. Early starting of treatment determines the probability of achievement higher osustain viral response.

COMPARATIVE STUDY OF FIB-4 INDEX AND TRANSIENT ELASTOGRAPHY AMONG PATIENTS WITH CHRONIC HEPATITIS C VIRUS INFECTION IN GEORGIA.

Liver biopsy remains the reference standard for fibrosis staging. However, it has several limitations, which have led to the development of non-invasive methods. We evaluated liver fibrosis severity among HCV infected patients by comparing transient elastography (TE) and FIB-4 index. Retrospective study was conducted. Clinical data for 750 patients were obtained. The mean age of the study population was 51 years; 595 (79.3%) were male and 155 (20.7%) were female. TE and tests on biological samples were performed within one-week timeframe. Additional analyses of prothrombin index, albumin concentration, splenomegaly on abdominal ultrasound and esophageal varices on upper gastrointestinal endoscopy were performed among selected patients. Comparable results were observed among 534 patients (71.2%). FIB-4<1.45 had a negative predictive value of 89% to exclude significant fibrosis and FIB-4>3.25 had a positive predictive value of 100 % to confirm the existence of significant fibrosis. Inconclusive FIB-4 score was obtained in 170 (22.7%) patients. Of them 127 (74.7%) had significant fibrosis (F3-F4) by TE. Discordant results (FIB-4 <1.45 and Liver Stiffness Measurement (LSM) >9.5 kpa) were observed in 46 (6.1%) of patients. Low prothrombin index, low albumin concentration, splenomegaly and esophageal varices were significantly (p<0.001) correlated with TE results. Discrepancy showing high FIB-4 score and low LSM was not observed in our cohort. There was a good correlation between TE and FIB-4 score. FIB-4 could rapidly replace expensive methods to assess liver fibrosis severity in some scenarios. However, our study demonstrated superiority of TE. LSM correlated better with indirect markers of significant fibrosis.

INCIDENCE OF TUBERCULOSIS AMONG HIV/HCV CO-INFECTED PATIENTS RECEIVING HEPATITIS C TREATMENT WITH PEGYLATED INTERFERON AND RIBAVIRIN IN GEORGIA.

Treatment of hepatitis C is necessary for ensuring higher life expectancy among HIV/HCV co-infected patients. However antiviral treatment for chronic HCV infection with Pegylated interferon (PEG-IFN) and Ribavirin (RBV) is associated with a variety of side effects. In Georgia up to 22% of HIV-infected patients were found to have active Tuberculosis (TB) and 22.4 to 32.6% had latent TB. The objective of this study was to describe the characteristics and clinical outcomes of tuberculosis in HIV/HCV co-infected patients receiving hepatitis C treatment with pegylated interferon and ribavirin and calculate incidence rate of TB. A retrospective study was conducted among HIV/HCV co-infected patients receiving antiviral treatment for chronic HCV infection at the Infectious Diseases, AIDS and Clinical Immunology Research Center, Tbilisi, Georgia from December 2011 to May, 2015. A total of 420 HIV/HCV co-infected patients received HCV therapy with PEG-IFN and RBV during study period. Six of 420 patients developed TB while receiving PEG IFN + RBV therapy. These patients were on Antiretroviral treatment. Baseline HIV RNA load was <34 copies/ml and CD4+ cell counts >350 cells/mm3. No opportunistic infections were observed in all cases. Three of 6 patients had a previous positive tuberculin skin test (TST) result and had completed isoniazid chemoprophylaxis several years before TB diagnosis. In 2 patients TST was not performed. Only one patient had experienced a previous episode of TB and had completed the anti-TB therapy 1 year before hepatitis C treatment. In all patients TB was diagnosed during the PEG IFN + RBV therapy. Hepatitis C treatment was immediately stopped in all patients. The incidence rate of TB was 1.4 cases per 100 person-years (95% CI=0.58-2.97). Our study emphasizes the necessity of screening for latent TB prior to the initiation of chronic hepatitis C treatment with PEG IFN and RBV.

MOLECULAR TRACING OF HETEROSEXUAL HIV-1 TRANSMISSION IN GEORGIA.

HIV epidemic in Georgia has entered a new phase with number of heterosexually acquired infections rising each year. Epidemiological data indicates that this switch in epidemic trends is largely due to HIV positive male IDUs transmitting the virus to their female sexual partners. However, no genetic studies confirming linkage between IDUs and their sex partners were done in Georgia before. The objective of our study was to investigate molecular epidemiology of HIV-1 transmission events between heterosexual couples. Viral genotypes were obtained from plasma specimens of 36 heterosexual HIV-1 positive antiretroviral treatment (ART) naive persons representing 18 epidemiologically linked transmission events were genotyped and phylogenetic analyses were done on HIV pol sequences. HIV infection among all women was attributed to heterosexual transmission from their partners. None of 18 women had history of IDU. Fourteen pairs had subtype A virus, three - subtype B and one - subtype G viruses. Phylogenetic analysis confirmed the existing epidemiological link in 16 pairs with bootstrap values ranging from 88% to 100%. Of these 16 events, viruses from 14 pairs had genetic distance less than 0.015.Mutation A62V was seen in samples from 5 pairs, of them samples from 4 pairs additionally had V77I mutation. All 5 pairs were infected with the subtype Avirus. Women, who are sexual partners of IDUs or other men with high risk heterosexual behaviors, are at increased risk of HIV acquisition. HIV epidemic in Georgia has not spread to general population and remains concentrated around key populations at risk. Our work confirms that female sexual partners can serve as a bridge between key affected populations and general community, such as heterosexually active adults. Therefore, prevention efforts targeting key populations at risk and their sexual partners need to be expanded to avoid the spread of the infection within specific communities and beyond.

Distinct drug resistance profile of HIV-1 subtype A strain circulating in Georgia.

Emergence of HIV-1 drug resistance limits effectiveness of antiretroviral therapy (ART). Since 2004 Georgia provides free ART to all patients in need. We aimed to evaluate drug resistance patterns of Georgian HIV-1 variants among patients with virologic failure. Study included adult HIV-1 patients, who experienced virologic failure and were found to carry drug resistant strains based on genotypic resistance testing in 2005-2013. HIV-1 pol gene sequences were examined for the presence of resistance-associated mutations. Stanford HIV Sequence Database was used for interpretation of resistance data. A total 193 patients were included in the study. Among them majority (86.5%) carried subtype A virus and nearly 80% were on Efavirenz-based regimen. The most common nucleoside reverse transcriptase inhibitor (NRTI) mutation was M184V - 86.0% (n=166). The most frequent non-nucleoside reverse transcriptase inhibitor (NNRTI) mutation was G190S, found in 105 (54.4%) of samples. Other significant NNRTI mutations included K101E (31.6%, n=61), K103N (30.1%, n=58) and Y181CI (26.9%, n=52). The prevalence of G190S was 62.3% in subtype A viruses compared to 3.8% in non-A variants (p<0.0001). Frequency of K101E was also significantly higher in subtype A (36.5% vs. 0%, p<0.0001). ). In 69 samples G190S co-occurred with either K101E or Y181C or with both: 39 genotypes G190S/K101E; 10 genotypes G190S/Y181CI and 20 genotypes G190S/K101E/Y181CI. High prevalence of G190S and K101 mutations suggests subtype A specific response to currently approved first-line NNRTIs. Frequent co-occurrence of G190S with Y181C and K101E may limit the use of novel generation NNRTIs in subtype A infected patients with previous exposure to this drug class.

Discrepancy between HCV structural and non structural genes in Georgian genotype two patients.

Correct identification of hepatitis C genotypes is an important diagnostic tool, which guarantees further selection of adequate treatment regimen and correct duration. Ideal approach for accurate genotyping is amplification of both structural and non structural parts of HCV genome. As different methods, which use either one or another region for HCV genotyping sometimes lead to indeterminate genotype and subtype results. Therefore, it is of importance to compare HCV genotyping results based on two different genomic regions. As part of this study, remnant 108 specimens, with previous history of successful genotype identification by 5'UTR/core Versant HCV genotyping kit, were retrospectively analyzed. "In house" HCV real time PCR based method that amplifies parts of NS5B region was used for this purpose. Based on our data, genotyping calls were concordant between genotype one and genotype three specimens group in both regions. However, discrepancy was evident among genotype 2 group. Of 25 specimens originally typed as genotype 2 in structural region, only 7 was confirmed in non structural region, remaining 18 specimens were typed as 1b. Therefore, the discordance rate between structural and non structural regions for genotyping call among genotype two was 72%. Our data showed highly discordant structural and non structural genome for genotype two identification in our specimens. We propose that this phenomenon might be due to the recombination event between genotype two and genotype one. Possible circulation of this strain in Georgia stresses the need for detailed sequencing and phylogenetic analyses of these specimens in both structural and non structural parts of HCV genome.

Occurrence of occult HCV infection among Hiv infected patients in Georgia.

Occult hepatitis C (OCI) infection has been known as detectable HCV-RNA in the liver or peripheral blood mononuclear cells (PBMCs) in the absence of detectable serum or plasma HCV-RNA. OCI has been detected among different patients groups worldwide, it has been found not only in chronic hepatitis patients of unknown origin, but also among several groups at risk for HCV infection (hemodialysis patients or family members of patients with occult HCV). This occult infection has been reported also in healthy populations without evidence of liver disease. Prevalence of occult Hepatitis C virus has not been investigated in Georgian population, where a rate of HCV infection is highest (6.7%) among Eastern European Countries. The aim of this study was to investigate the occurrence of occult HCV infection among HIV infected individuals in Georgia. As a pilot study, we have selected three groups of HIV infected patients for analyses: Group 1- HIV infected patients without evidence of liver disease (n=98), group 2- HIV infected patients with cryptogenic liver disease (n=34) and group 3- HIV/HBV co infected patients (n=29). HCV RNA was tested in PBMCs samples by real-time polymerase chain reaction. HCV genotyping was performed by Line-probe assay based on reverse-hybridization technology. Liver fibrosis was evaluated by transient elastography (FibroScan®). HCV-RNA was detected in PBMCs specimens among 2 (2%) subjects from group 1, 4 (12%) subjects from group 2, and 9 (31%) subjects from group 3. HCV genotypes were determined for 14 of 15 OCI subjects resulting following genotype distribution: 6 (46%) - 1b, 3 (23%) - 2a/2c and 5 (38%) - 3a. One samples failed to be genotyped due to extremely low HCV viral load. Our data revealed the occurrence of occult HCV infection in HIV infected patients. No single HCV genotype was predominant in the present study. Liver fibrosis was found more frequently and the fibrosis score was significantly higher in OCI patients versus negative ones, suggesting that undiagnosed OCI might impact on the liver damage. The study demonstrated that testing only for HCV antibody fails to identify the true prevalence of HCV co-infection among HIV infected patients. We propose that in the absence of liver biopsy specimens, analysis of PBMC sample for HCV-RNA would be informative for detection of occult HCV.

[Comparative diagnostic value of Helicobacter pylori infection testing methods].

In 213 patients with gastric and duodenal pathology, including received surgery, comparative estimation of results of Helicobacter pylori (Hp) infection testing with invasive and noninvasive methods, were performed. Material for invasive endoscopic biopsy test (EBT), including rapid urease test (RUT) for rapid Hp identification by determination of urease activity, smear cytology and histology was extracted on endoscopy or intraoperationally. RUT was carried out with the help of URE-HP test kit. Serological test for Hp antibodies IgG and IgA class was performed by IFA using kit ELISA. 13С urea breath test (UBT) was made by determination of 13/12CO2 in breath samples on infrared spectroscope. Based on 5 different methods of Hp infection testing Hp positivity in 172 (80,8%) and Hp negativity in 41 (19,2%) from 213 examined patients was revealed. 13С-UBT revealed the highest diagnostic value (accuracy-97,5%, sensibility-97,0%, specificity-100%) in Hp infection diagnostics. In treatment efficiency control this parameters of 13С-UBT are also much high (96,7%, 90,0% and 100% respectively). In spite of high sensitivity of serological test (100%), it had comparative low specificity (71,0%) with high probability of false positive results in treated patients (antibodies titer to Hp after eradication retains for a long time). Thought, this test may be successfully used only in primary patients and in epidemiological studies. Among three methods of EBT, Hp infection detection with RUT revealed the best results (accuracy-94,8%, sensibility-95,0%, specificity-100%). Correlation of RUT and UBT results and much higher diagnostic value of UBT, necessitate RUT with histological study of stomach body mucosa to perform in patients over 45 year, with prolonged anamnesis and dangerous symptoms of disease. In Hp positive patients correlation of index DOB‰ of breath test with results of RUT was revealed. This can serve the index DOB‰ of 13С-UBT as a marker of Hp infection rate. The (accuracy, sensibility and specificity of breath test with local made 13С-urea (98,7%, 98,5% and 100% respectively) are the same as those for BT with standard 13С-urea (96,7%, 96,2% and 100% respectively). Above mentioned, 13С-UBT advantages (noninvasiveness, simplicity, rapidity, safety) and high diagnostic value exceeding same of other applied tests, give us opportunity to offer 13С-UBT as screening method of Hp infection diagnosis and a method of choise in control of Hp infection treatment efficiency.

High seroprevalence of Chlamydia trachomatis in newly diagnosed human immunodeficiency virus patients in georgia.

Due to the shared routes of transmission, co-infection with Human Immunodeficiency Virus (HIV) and other sexually transmitted infections (STIs) is common. There is strong evidence of bidirectional interactions between HIV and ulcerative STIs. Recent studies have also shown importance of non-ulcerative inflammatory STIs in the acquisition of HIV. The incidence of HIV and Chlamydia in Georgia has risen every year. We explored the extent of the problem of co-infection with C. trachomatis in HIV patients in the country. Study included 234 consecutive patients diagnosed with HIV from September 2008 through May 2009. Of them, approximately two-thirds were male 162 (69.23%), up to 44% (102) of patients had more than one lifetime sexual partner and one fifth of patients reported prior history of STIs. The seroprevalence of C. trachomatis in our study was 23.93% (95% CI: 18.61%-29.92%). In multivariate analysis the strongest predictors of C. trachomatis infection were history of STI (PR 1.94, 95% CI: 1.22-3.07) and female gender (PR 1.79, 95% CI: 1.11-2.87), while younger age and not being in marriage showed borderline significance. Findings of our study have important public health and clinical implications. Data suggest that STIs may play important role in increasing heterosexual transmission of HIV in Georgia. Efforts should be made to expand HIV screening programs. Further research is needed to better understand the role of inflammatory STIs in spreading HIV.

HLA-B*5701 genetic screening prior to abacavir prescription in Georgia.

A hypersensitivity reaction to abacavir develops in approximately 2-8% of HIV patients receiving this drug and is strongly associated with presence of the human leukocyte antigen (HLA)-B*5701. Screening for HLA-B*5701 reduces the risk of developing an abacavir hypersensitivity reaction. The carriage rate of HLA-B*5701 has not been studied in Georgia before 2009. Objective of the study was to determine HLA-B*5701 prevalence in HIV-infected patients in Georgia. One hundred and sixty HIV positive patients attending Georgian Infectious Diseases, AIDS and Clinical Immunology Research Center in 2009 were recruited for the study. None of the patients had previously been treated with abacavir. Blood samples were collected and screened for HLA-B*5701 prior to abacavir prescription. Of 160 patients recruited 9 tested HLA B*5701 positive - 5.6% (95% CI: 2.6-10.4%). Of these nine patients 7 were males (male prevalence: 6.5%, 95% CI: 2.6-12.9 %) and 2 females (female prevalence: 4.8%, 95% CI: 0.6-16.2%). The first prospective study of HLA-B*5701 prevalence in Georgia show similar results to the results of other studies. Abacavir still remains one of the key drugs of antiretroviral regimens in Georgia and other countries. Therefore, prospective HLA-B*5701 screening should be implemented in all settings where abacavir is widely used to guide selection of ART regimens and to reduce the risk of potentially life threatening hypersensitivity reaction.

Safety and efficacy of systematic administration of Filgrastim to prevent neutropenia and infections in patient with hepatitis C.

The aim of 36 months follow up study was to assess the safety and efficacy of Filgrastim (Neupogen) for preventing neutropenia and bacterial infection during combination therapy of chronic HCV infection with pegilated interferon alfa and ribavirin. Study enrolled 64 patients with chronic active hepatitis C, aged 20-65. Among them 49 were male and 15 female). Among 64 patients: 5 patients had HCV genotype 1a, 24 patients HCV genotype 1b, 17 patients HCV genotype 2a/2c and 18 patients HCV genotype 3a. Treatment regimen for chronic hepatitis C patients was as follows: Pegylated interferon alfa 2a (Pegasys) 180 micro kg or alfa 2b (PegIntron) 1,5 micro g/kg. and ribavirin (RBV). RBV daily dose was adjusted by body weight- 1000/1200 mg. Treatment duration was 48 weeks for HCV genotype 1 patient and 24 weeks for HCV non 1 genotype accordingly. The patients were divided into two groups: 29 patients (1st group) besides combination antiviral therapy (pegilated interferon alfa plus ribavirin) systematically received Filgrastim and other 35 patients (2nd group) - same antiviral therapy without administration of Filgrastim. Selection of patients was performed by computerized randomization method. HCV antibodies were detected by ELISA and RIBA. HCV RNA by Real time PCR. HCV genotype- by Inno-Lipa. Among 2nd group 35 patients (without Filgrastim administration) during antiviral therapy 8 patients (22.8%) developed different bacterial infections.(3 patients- urinary tract infection, 2 patients- pneumonia, 1 patient- bronchitis, 1 patients - sinusitis and 1 patient-gingivitis/stomatitis). 7 patients required interferon dose modification (dose reduction) and in 5 patients treatment stopped due to severe neutropenia. Among 1st group patients (with filgrastim administration) only one patient developed bacterial infection (urinary tract infection). None of patients, due to neutropenia, required neither stoppage of therapy, nor interferon dose reduction. The quality of life of 1st group patients was better in comparison of 2nd group patients. Filgrastim was safe and effective for prevention neutropenia and bacterial infections in Hepatitis C patients with Peg-INF/RBV combination antiviral therapy. Filgrastim was well tolerated by patients. It gives possibility to maintain interferon dose during treatment period and significantly improves the patient's quality of live.

Re-treatment of patients with hepatitis C who failed to respond (nonresponders) to previous treatment.

The aim of four-year follow up study was evaluation of re-treatment efficacy of antiviral therapy in patients with hepatitis C who failed to respond (non responders) to previous therapy. Study enrolled 29 patients, aged 21-59 with HCV infection (15 had HCV genotype 1, and 14 had HCV non-genotype1), who previously were treated with unmodified interferon alfa (conventional interferon) 2a or 2b 5 MIU TIW plus ribavirin (1000-1200 mg/day) and who failed under this therapy. Study subjects were randomized into two groups: in group I were included 17 patients--relapsers (patient in whom HCV RNA becomes undetectable on treatment and is undetectable at the end of therapy, but is detected again after discontinuation of treatment). Group II was composed of 12 patients: 4 were non responders (patient in whom HCV RNA levels remain stable on treatment), 4--partial responders (HCV RNA levels decline by >2 logs, but never become undetectable during treatment) and 4--breakthrough non responders (HCV RNA become undetectable during treatment, but before-treatment termination again become detectable). The diagnosis of HCV infection was made based on detection of HCV antibodies by ELISA and confirmed by RIBA. Detection of HCV RNA (qualitative) and HCV RNA Viral load--by Real time PCR technique (COBAS TaqMan Test). HCV genotypes were detected by INNO-Lipa method. In group I--rapid virological response (RVR) was observed in 10 (58%) patients, early viral response (EVR) in 12 patients (70%). Among them 9 (52%) patients remained HCV RNA undetectable by the end of treatment. After 6 months sustained viral response (SVR) was received in 7 (41%) patients from group I. In group II--RVR was observed in 5 (41%), EVR in 6 (50%) patients. Among them 5 (41%) patients remained HCV RNA undetectable by the end of treatment. After 6 months Sustained Viral Response was received in 3 (25%) patients. Re-treatment with pegylated interferon and ribavirin in patients with hepatitis C who failed to responds to previous treatment was effective in relapsers. Re-treatment in non responders, partial responders and breakthrough non responders was less effective (especially in non responders). Re-treatment effectiveness was higher in HCV genotype non 1 patients in comparison with HCV genotype 1. Thus re-treatment will be considered for relapsers. For making decision on re-treatment for other nonresponders, severity of disease (advance disease) should be considered.

Prevailing HCV genotypes and subtypes among hiv infected patients in Georgia.

Recent analysis of antiretroviral treatment (ART) program data in Georgia showed that end-stage liver disease was a leading cause of death among HIV/HCV co infected patients in 2005. The objective of this retrospective study was to study prevailing genotypes and subtypes of HCV virus in a cohort of HIV infected patients. The investigation revealed that of 1490 patients, 879 (59%) were hepatitis C antibody positive. Detectable HCV RNA was found among 91% of patients. Median liver HCV RNA level was higher than among mono-infected patients. The most prevalent genotypes were genotype 1 (41.6%), followed by genotype 3 (34.7%) and genotype 2 (17.6%), inter (mix) genotype recombinants were found among 5.8 % of patients. The genotype distribution in our study is slightly different from what was seen in Georgia in 2000. The differences of prevailing HCV genotypes among general population and HIV co infected group was probably attributed to the different methods for sample selection used within our study or possible influence of diverse transmission networks among HIV infected group. Another explanation can be the possible shift from predominance of genotype 1 to non 1 genotypes. The higher number intergenotype recombinant forms might be the result of continues parenteral exposure to different HCV genotypes during drug injection paraphernalia. Our study demonstrated high prevalence of HCV infection among HIV-infected patients and revealed 1b as predominant genotype. IDUs were less likely to spontaneously clear the virus than homosexual man and heterosexually infected woman. A greater HCV RNA levels were associated with a greater chance to be infected with HCV genotypes 1. Possible shift from predominance of genotype 1 to non 1 genotypes can be of option. This shift may have a major and beneficial impact on treatment schedules and costs. The higher number intergenotype recombinant forms might be the results of continues parenteral exposure to different HCV genotypes during drug injection paraphernalia. Study results highlights and strengthens the need for careful follow-up of HCV/HIV co infected patients, effective management and therapies against HCV in order to reduce liver related death rates in patients on ART.

Antiretroviral treatment in Georgia.

HIV infection is the major public health, social and economic problem in Georgia. The aim of this study is to evaluate effectiveness of ARV treatment system in Georgia. Study included 1052 people living with HIV/AIDS in Georgia registered at Infectious Disease, AIDS and Clinical Immunology Research Center since 2004. To ensure universal access to ARV therapy all HIV/AIDS individuals included in the study were investigated by special algorithm, all identified patients requiring ARV therapy were offered treatment and monitored during therapy on treatment effectiveness and side effects. Detection of HIV antibodies was performed by ELISA with further confirmation by Western Blot Assay. HIV-1 RNA in plasma was measured by quantitative Polymerase Chain Reaction. For determination of percentages and absolute count of T lymphocyte subpopulations single-platform immunophenotyping technique using the Becton-Dickinson FACSCalibur flow cytometer was applied. For resistance testing TRUGENE HIV-1 Genotyping Kit with the OpenGene DNA Sequencing System (Siemens) was used. Treatment was offered to 595 HIV/AIDS patients. 594 patients started treatment, 1 patient refused. Out of treated 594 HIV/AIDS patients 22 patients discontinued, 111 patients died and 461 patients are currently on ARV treatment. Out of treated patients 406 adults and 21 children are receiving first-line treatment, 31 adults and 2 children are on second-line treatment and 1 adult is receiving salvage regimen. Treatment failure was defined in 55 cases. Among them immunological failure was observed in 7 cases, clinical failure in 1 case and virologic failure in 47 cases. Prevalence of drug resistance among virologic failure cases accounted for 72% and inadequate adherence for 28% cases. Majority of death cases among ARV treated patients was due to non-AIDS related or incurable conditions, while deaths due to AIDS related conditions mainly were associated to the delayed referral of patients in already advanced stage of disease. It's worth to mention that highest number of death cases was due to liver failure in HIV/HCV and/or HBV co-infected patients.

Successful application of laboratory tools for the detection of HIV drug resistance in routine clinical care in Georgia.

Since 2004, Georgia the first among Eastern European countries ensured universal access to highly active antiretroviral therapy (HAART). Laboratory monitoring of HAART using CD4 count, viral load (VL) and HIV genotypic resistance testing was carried out in according with National HIV/AIDS Treatment Guidelines. Georgia the first among former Soviet Union countries implemented HIV genotypic resistance testing in HIV clinical care. The present paper reports on successful application of laboratory tools in routine clinical care for the early detection of HIV drug resistance. For genotypic resistance testing the TruGene HIV-1 Genotyping Kit (Bayer HealthCare LLC, Tarrytown, NY) was used according to manufacturer's instructions. Analysis included 45 patients with virologic failure. Of them 34 (75.5%) had at least one resistant mutation. Dual-class drug resistance was found in 30 (66.7%) patients. One (2.2%) patient carried triple-class resistance mutations. Median number of resistant mutations was 2. Most commonly detected NRTI mutation was M184/V/I (68.9%). G190S/A was the most frequent NNRTI mutation (42.2%), followed by K103N (28.9%). All patients with drug resistance mutations were switched to a second line regimens. Analysis of virologic and immunological outcomes among 23 patients who had at least two follow-up measurements of CD4 and VL after resistance test, showed statistically significant decrease in VL by 2.5 log(10) and mean gain of 181 cells/mm(3) in CD4 count by the last available measurement. Routine monitoring of VL and subsequent use of HIV drug resistance testing allowed for early identification of HIV drug resistance, reducing the opportunity for mutations to accumulate. Routine use of sophisticated laboratory methods for HAART monitoring has beneficial impact on clinical outcomes and should be used as part of the strategy to combat resistance.

Assessment of liver fibrosis and cirrhosis by transient elastography among patients with chronic HBV and HCV infection in Georgia.

The aim of the study was to evaluate liver fibrosis (LF) and cirrhosis using Transient elastography (TE) using Fibroscan in patients with chronic HBV and HCV infection. The device evaluates LF by measurement of liver stiffness (LS). 525 patients with chronic HCV infection and 105 patients with chronic HBV infection were included in the study. These patients were investigated at the Georgian-French joint hepatology clinic "Hepa" from November 2007, till November 2008. Among investigated HBV infected 105 patients 65 (61.9%) had no fibrosis (LS<5.5 kpa), 23 (21.9%) had mild fibrosis (LS-5.5-8.0 kpa), 9 (8.6%) had severe fibrosis (LS-8.0-14.0 kpa) and 8 (7.6%) had liver cirrhosis (LS>14.0 kpa). Among investigated HCV infected 525 patients 200 (38.1%) had no fibrosis (LS<5.5 kpa), 139 (26.5%) patients had mild fibrosis (LS-5.5-8.0 kpa), 87 (16.5%) patients had severe fibrosis (LS-8.0-14.0 kpa) and 99 (18.9%) patients had liver cirrhosis. It is concluded that transient elastography (TE) using Fibroscan is simple, non-invasive, reliable and easily reproducible method for assessing liver fibrosis and cirrhosis in patients with chronic HBV and HCV infection. TE is characterized with an excellent accuracy. TE results are well correlated with the clinical signs as well as with the results of laboratory and instrumental investigations. Fibrosis stages by Metavir measured using Fibroscan well corresponds with the liver biopsy results. Considering the high prevalence of fibrosis and cirrhosis among patients with chronic HBV and HCV infection, TE is a very valuable method for detecting early stages of fibrosis allowing to avoid the progression of liver damage, as well as end-stage liver disease. TE is easy to perform and therefore allows regular follow-up of the course of LF.

Acute/recent HCV infection. Clinical course, viral replication kunetic and disease outcome.

The aim of the study was to reveal and investigate acute/recent HCV infection at the very early stage in seronegative blood donors and seronegative Injecting Drug Users (IDUs) and to assess clinical laboratory variants of infection, viral replication kinetic, disease outcome, host and viral characteristics. Two groups of patients were included in this study. The first group consisted of ELISA negative 7000 blood donors; the second group included 3000 Injecting Drug Users (IDUs). All patients were investigated on HCV RNA by qualitative PCR using mini pool method. A pool of 6 was applied for blood donors' and a pool of 5 for IDUs. PCR negative pools were excluded from the study, while PCR positives were examined on individual samples. Anti-HCV was detected by ELISA and RIBA. Detection HCV RNA was performed by Real time PCR technique using COBAS TaqMan Test. HCV genotyping--by INNO-Lipa. HLA typing--by Sequence Specific Primer Amplification (SSP). 16 patients with acute/recent HCV were revealed: 7 from blood donors, 9 from IDUs. Among them: 4 were symptomatics and 12 asymptomatics. Out of 4 symptomatics 3 were with jaundice. Among 12 asymptomatics: 8 had elevated ALT; 2 neither elevated ALT nor symptoms but developed anti-HCV; 2 were with normal ALT and without further anti-HCV seroconversion. Among 16 subjects: 9 had genotype -1b, 1--genotype 1a, 3--genotype 2a/2c and 3--genotype 3a. Out of 16 cases 4 cleared the virus; 12 developed chronic infection. Spontaneous clearance (recovery from the disease) was observed in 2 out of 4 symptomatic patients and only in 2 patients out of 12 asymptomatics. In all patients viremia increased rapidly and reached a peak by week 4. Viral titer was remarkably stable for the next three weeks, followed by two or three fold decrease by week 9. After week 10 the viremia rapidly decreased: 4 or 5 logs by week 12 and it became either undetectable by weeks 16-18 (viral clearance), or virus was not eliminated and viral titer persisted in all follow up period (chronic infection). HLA DRB1 1101, DQB1 0301 and DRB1 1301/DQA1 0103 alleles were associated with clearance of HCV whereas DRB1 0301 was associated with chronic infection. Prevalence of HCV among seronegative blood donors was 0.1% and among IDUs 0.3%. Among acute/recent HCV infected patients rate of chronicity was 75% (50% in symptomatics and 83% in asymptomatics). Rate of recovery was 50% in symptomatic patients and about 16% in asymptomatics. Acute/recent HCV infection might have following clinical laboratory forms: symptomatic disease with or without jaundice, asymptomatic with or without elevated ALT, but with further anti-HCV seroconversion. It remains unclear whether enigmatic form of disease--acute/recent HCV infection without further seroconversion exists or not.

Prevalence of hepatitis B and C among HIV positive patients in Georgia and its associated risk factors.

The aim of the study was to determine the prevalence of hepatitis B virus (HBV) and hepatitis C virus (HCV) co-infection among HIV positive patients, to identify most relevant risk factors of co-infection and develop preventive interventions. Study participants were voluntary individuals 18 years of age or older recruited from AIDS Center VCT unit in Tbilisi, Georgia. Eligibility criteria of participants were: HIV positive result confirmed by western blot; age; and voluntary participation. Total 175 patients undergo interview with specially designed questionnaires. Most of the participants were male (71.4%), age range of HIV positives varied from 20 to 77 years old. Prevalence of HCV among HIV positive patients is high. Almost half (48.57%) HIV positive patients are co-infected with HCV. Men were more likely than women co-infected with HCV (60.80% and 18% accordingly). Major risk factor of male co-infection was related to drug use, needle and injection equipment sharing. Prevalence of HCV among injecting drug users was (73.40%). Drug users had 3.25 times more risk (PR 3.25; 95%CI; CL--1.89-5.26; p<0.01) to be infected with HCV compare non IDUs. Prevalence of being infected with HBV (Anti-HBc) among HIV positives was 43.42% (76/175) and the prevalence of Chronic HBV (HBsAg positive) was 6.86% (12/175). Prevalence rate of HBsAg among IDUs was 8.51% and among non IDU participants 5.26%. Triple infection (HIV, Hepatitis C and chronic form of Hepatitis B--HBsAg) was among 9 patients (5.14%). Infections were associated with injection drug use (88.88%) and mostly were related to share of needles/syringes and other injecting medical equipment. Transmission of HBV and HCV by sexual contact was not observed among those 9 participants. High risk behavior among HIV positive participants mostly related to drug use and unprotected sex with non regular partners. Other risk factors for Hepatitis transmission were associated with invasive medical manipulations, blood transfusion, surgery, abortions and etc. None of cases of HIV, or Hepatitis (B, C) transmission through medical manipulations can be documentary proved based on those research data.

Prevalence of HCV and genotypes distribution in general population of Georgia.

The aim of four years study was to determine the prevalence of HCV infection in the general population of Georgia and to assess HCV genotypes spread among them. For performing the planned investigation a cross-sectional study design was applied. Study subjects were Tbilisi residents selected through multiple clusters sampling technique application. Tbilisi is divided into ten districts. 2000 persons from the general adult population of Tbilisi, Georgia were enrolled in the study. The multi-stage cluster sampling method was applied for selection of study subjects. Districts of Tbilisi were considered as sub-populations. At the first stage the number of Population distribution by each districts were calculated. Number of study subjects by each districts were determined proportionally population. A list of study participants were obtained from primary sampling units--policlinics (Primary Health Care Units--PHCUs) in all ten districts of Tbilisi. A fieldwork was arranged for interviewing of selected individuals--Approaching the study subjects in households, Face-to-face interview which included signing of the letter of consent by study subject and administering the questionnaires by an interviewer. In totally 2000 persons were investigated and accordingly 2000 blood samples were tested by ELISA for detection of HCV antibodies. 138 out of 2000 (6.9%) samples were found ELISA positive. 138 ELISA positive samples were tested with more specific Recombinant Immunoblot Assay (RIBA) for confirmation. 134 out of 138 (6.7%) of investigated samples were confirmed by RIBA as positives. In this survey, we found that 134 (6.7%) of the 2,000 surveyed individuals were HCV seropositive. According our study, which was based on very strict epidemiological design, we concluded that prevalence of HCV in General population of Georgia is 6.7%. The part of our investigation was to assess HCV genotypes distribution among general population of Georgia. Based on our results the following distributions of HCV genotypes were found: HCV 1b--59%, HCV 3a--27%, HCV 2a/2c--11%, HCV 1a--3%. Our study found high prevalence of HCV among general population of Georgia. Besides, these surveys found an extensive spread of HCV 1b genotype. The profile of HCV genotypes distribution in general population of Georgia was similar to that of USA and Russia and different compared to Asia, Africa and most of European countries. Unfortunately the HCV genotype 1b is less sensitive to current treatment regimens. The treatment effectiveness in patients with HCV genotype 1b is about 45% in comparison to 80% for non-1 genotypes.

IFN/RBV treatment induced neutropenia and its correction with neupogen in patients with hepatitis C.

The aim of the study was to observe the frequency of neutropenia during Pegylated Interferon/Ribavirin therapy in patient with chronic hepatitis C; to compare the efficacy of two strategies of management of neutropenia--with Interferon dose modification and with Neupogen administration; to compare the effectiveness rate of sustained viral response (SVR) in patients with Pegylated Interferon dose modification and in patients treated by using granulocyte colony-stimulating factor G-CSF-filgrastim. (Neupogen). Study enrolled 47 patients with chronic active hepatitis C, aged 23-64. (38 male and 9 female). All patients had HCV genotype 1b. Significant neurtopenia (ANC<750 mm3) and severe neurtopenia (ANC<500 mm3) developed in 41 of 47 patients (87%). 41 patients with neurtopenia were randomized into two groups. The first group--22 patients who received granulocyte colony-stimulating factor (G-CSF, or filgrastim) 300 mcg s/c weekly for correction of neutropenia and the second group--19 patients treated either with Interferon dose reduction or temporarily inhibit of Interferon treatment. In all 22 patients of the first group neutropenia was normalized without reduction and/or inhibit of Pegylated interferon. Neupogen was well tolerated and in all 22 patients the improvement of quality of life (QOL) was observed. It was concluded that dose reduction or temporary inhibit of Pegylated Interferon in the second group negatively acts on antiviral treatment response in patients with HCV genotype 1. In patients with PEG-IFN/RBV therapy Neupogen effectively manages neutropenia and gives opportunity to maintain interferon dose (without reduction). Neupogen has the potential to improve adherence rates, which may in turn improve SVR.