[The diagnosis of idiopathic epilepsy in children based on the algorithm of molecular-genetic studies]. / Diagnostika idiopaticheskikh form epilepsii u detei na osnovanii algoritma molekulyarno-geneticheskogo issledovaniya.

AIM: To study mutations and polymorphisms in the sodium channels genes, determining the development of idiopathic epilepsy (IE). MATERIAL AND METHODS: The study of SCN1A gene by direct Sanger sequencing in 53 patients and targeted resequencing of the regions of 34 genes in 40 patients with different clinical forms of IE was performed. RESULTS: Seven mutations (c.3022G>T, c.3637C>T, c.1144G>T, c.80G>C, c.1603C>T, c.2427G>A and c.1131A>C) were detected among 53 patients by direct Sanger sequencing of SCN1A gene. The mutations of SCN1A gene (2 - nonsense mutation, 5 - missense mutation) were identified in 7/40 (17.5%) patients with epilepsy using high-performance sequencing, Mutations in sodium channel genes encoding other subunits: SCN1B, SCN2A, SCN9A were identified in 6 patients. CONCLUSION: As epileptic encephalopathy is polygenic, it is important to conduct genetic testing of more genes (primarily sodium channel genes - SCN1B, SCN2A, SCN9A etc.) using special gene panels to find the molecular defect in DNA.

[Prevalence of hepatitis E markers in children].

AIM: Frequency of detection determination for past and current hepatitis E virus (HEV) infection markers in children with immune suppression, as well as children with normal immune status. MATERIALS AND METHODS: The presence of HEV markers (anti-HEV IgG and IgM, HEV RNA) was studied in 609 sera samples of children with neurologic pathologies, 87 samples--from children with immune deficiencies, as well as 3122 samples from conditionally healthy children of 6 regions of Russia. The children were divided into 5 age groups. Anti-HEV IgG and IgM determination was carried out in EIA, HEV RNA--by RT-PCR. RESULTS: The frequency of detection of anamnestic anti-HEV IgG turned out to be significantly higher among immune-compromised. children compared with healthy children (5.7% against 1.4%, p < 0.05). Anti-HEV IgM, that testify to current or recent infection, were also detected significantly more frequently among children with immune-suppression (1.1-1.6%) compared with healthy children (0.25%, p < 0.05). HEV RNA was detected in 1 child with the absence of anti-HEV IgM and IgG. Nucleotide sequence analysis of HEV confirmed membership of this isolate in genotype 3, that is prevalent in non-endemic territories. CONCLUSION: The data obtained have demonstrated, that HEV-infection is prevalent among children in Russia and its course is, probably, asymptomatic in most cases. Immune suppression is a factor of increased risk of infection of children with HEV.

[FAMILIAL CLUSTERS OF HEPATITIS DELTA IN ENDEMIC REGION (REPUBLIC TYVA)].

AIM: Clinical and virological characteristic of hepatitis delta familial clusters in region of Russia that is endemic for this infection (Republic Tuva). MATERIALS AND METHODS: Total 383 patients with HBV/HDV coinfection and their family members (3 generations) were followed. Serum samples for HDV and HBV markers testing were available for 42 patients from 18 families. HBsAg, anti-HBc, HBeAg, anti-HBe and anti-HDV were tested using commercial ELISA tests; HDV RNA and HBV DNA were tested using in house nested PCR tests. RESULTS: 30 family (63 people) clusters were identified, in which close living persons have been infected with HDV and HBV. The biological material for determining of HDV and HBV markers has been available from 18 families (42 people belonging to 1-3 generations (parents and children, husband and wife, brother and sister). The mean age was 35 ± 14 years (10-58 years). Chronic hepatitis (CH) was in 30 (71.4%) patients, liver cirrhosis (LC)--in 10 (23.8%) and HCC was developed in 2 (4.8%) person on the background of long infections. The incidence of HBeAg was 14.3% (6/42), HBV DNA--19% (8/42); HDV RNA--35.7% (15/42). In 2 cases (mother) replication markers of both viruses were found, it contributed to the increased risk of infection in children. So HDV RNA was detected in the blood serum of their daughters (15 and 17), that does not exclude the possibility of vertical transmission. An illustration describes four families. CONCLUSION: The epidemic process of delta infection in the Republic of Tyva is characterized by intrafamilial infection of HBV and HDV.

[GENUS ANELLOVIRIDAE VIRUSES IN CHRONIC LIVER DISEASE].

AIM: Viruses from genus Anelloviridae (TTV, TTMDV and TTMV) are small DNA viruses that are widespread in human popu- lation. Data on tissue tropism, cell localization and morphometry of anelloviruses are scarce. The purpose of this study was to determine the prevalence of TTV, TTMDV and TTMV in persons with liver disease and in healthy individuals, as well as electron-microscopic verification of Anelloviridae species. METHODS: Detection of anelloviral DNA was performed in serum samples from 203 patients with liver diseases of various etiology and 115 voluntary blood donors using PCR with primers allowing to differentiate TTV, TTMDV TTMV based on the length of amplified fragment. Histopathological and electron microscopic studies were performed for liver biopsy specimens from 203 patients with liver disease. RESULTS: High prevalence (70-90%) of all three anelloviruses in healthy individuals and patients with liver disease was demonstrated, with high frequency of triple TTV, TTMDV and TTMV infection (52.2-55.7%). Electron-microscopic study of liver biopsy specimens from TTMDV monoinfected patients gave a submicroscopic image of TTMDV virions with diameter 35.86 ± 2.04 nm. Electron microscopic studies confirmed the nature of liver damage in TTMDV monoinfection: accumulation of virus in the hepatocytes, significant cyropathy with enlightenment matrix of the cytoplasm and reduction of intracellula organelles involved in protein synthesis, portal and perivascular perisinusoidal fibrosis. TTV, TTMDV and TTMV virions were dentified in hepatocytes, confirming these viruses to be hepatotropic. CONCLUSION: Our results demonstrate that anelloviruses are lymphotropic viruses, individual genotypes of those might be hepatotropic and pathogenic to liver.

[Viral hepatitis delta. Is there the delta infection problem in the Russian Federation?].

Hepatitis delta (HD) is characterized by rapid progression to fibrosis, and development of hepatocellular carcinoma, and a high mortality rate. The article presents data on the epidemiology, diagnosis, treatment of HD. The views of the epidemiological, clinical and virological characteristics of HD-infection among population of the Russian Federation (RF) are limited due to absence of official HD registration and detection of antibodies to the HD virus (anti-HDV) in HBsAg-positive individuals. However, some areas of the country are characterized by a high HDV circulation (Republic Tyva (RT) - 46,5%, Republic Sakha (Yakutia) - 12,5%) according to our studies conducted in 6 regions of Russia. Clinical-epidemiological situation of HDV infection in RT can be considered as a model to create a program of optimize diagnosis, prevention and treatment of HDV-infection in the Russian Federation.

[The panels of serums, containing various subtypes and mutant forms of HBsAg, to evaluate the diagnostics sensitivity of kits oa reagents detecting HBsAg].

The detection of HBsAg in blood serum using immune-enzyme analysis techniques decisively matters both for diagnostics of acute and chronic hepatitis B and screening of donor's blood and its components, controls of persons from risk groups of hepatitis B injection. The making of panels containing wide specter of samples of blood serums with sero-variants and mutant forms of surface antigen of hepatitis B virus widespread on the territory of the Russian Federation is necessary to control analytic and diagnostic sensitivity of test systems for detecting HBsAg. The testing of reagents kits to detect HBsAg using twi panels containing recombinant and native variants of HBsAg, demonstrated that these kits enable to detect various sero-vatriants of HBsAg (ayw2, adw2, ayw3varA, ayw3varB, adrq-) in concentration 0.1-0.01 IU/l and the so called elusive mutant forms of HBsAg of recombinant and native origin (G145R, Q129R, Q129H, Q129L, T143K, T126N, T126S, D144A, M133L, K141E and P142S). At that, the sensitivity can differ during the detection of native and recombinant mutant forms. The results testify the importance of using the panels both with recombinant and native samples containing the mutant forms of HBsAg in evaluation of sensitivity of reagents kits.

[Evaluation of the effect of mass vaccination against hepatitis B in the Russian Federation].

AIM: Evaluate the effect of mass vaccination against hepatitis B implemented within the framework of national project Health on the rate of development of unfavorable outcomes of infection and on average lifetime of the population of Russia. MATERIALS AND METHODS: Hepatitis B markers (HBsAg, anti-HBc, HBeAg, anti-HBe) were determined by enzyme immunoassayin 6211 blood serum samples from conditionally healthy population of 6 regions of the Russian Federation. Analysis of damage to health of the population caused by hepatitis B and favorable effect of mass vaccination against hepatitis B was performed by using mathematical model by Goldstein S.T. et al. (2005), that used results of detection of hepatitis B markers in discrete groups (women of childbearing age, children up to 5 years of age, adults aged 30 years and older). A formula developed by us was used to evaluate the effect of vaccination on the average lifespan of the population of the Russian Federation. RESULTS: The rate of HBsAg detection in the conditionally healthy population of the Tuva Republic was significantly higher (8.1%) than in other studied regions of the Russian Federation (p < 0.05): in Moscow Region--1.6%, in Rostov Region--1.6%, in Sverdlovsk Region--1.2%, Khabarosvk Region--2.0%, in Sakha (Yakutia) Republic--2.5%. The rate of detection of anti-HBc was significantly higher in the Tuva Republic and Sakha (Yakutia) Republic (46.2% and 42.3%, respectively); in other regions: Moscow Region--13.6%, Rostov Region--18.9%, Sverdlovsk Region--17.5%, Khabarovsk Region--21.0%. HBeAg in women of childbearing age was detected only in the Tuva Republic (0.4%). The mathematical analysis performed demonstrated that the level of coverage of vaccination of newborn children that in the studied regions is 96.1-99.6% may lead to the reduction of morbidity and mortality from hepatitis B and its outcomes by 91-95% that exceeds the similar parameter obtained by using our model, described in the literature for Africa, Asia and Europe. The calculated level of decrease of morbidity and mortality from hepatitis B and its outcomes may lead to the increase of average lifespan of the population of the Russian Federation by 1.44 - 4.08 months. CONCLUSION: The strategy of vaccination of newborn children against hepatitis B employed in the Russian Federation significantly reduces the risk of this disease and development of unfavorable outcomes of the disease especially in hyper endemic regions and promotes the increase of average lifespan of the population of Russia.

[Epidemiological and molecular biological analysis of causes of rise of hepatitis A incidence in Republic of Tuva in 2008].

AIM: Analysis of hepatitis A (HA) incidence in Tyva Republic in 2008 exploiting traditional epidemiological and molecular methods. MATERIALS AND METHODS: Epidemiological analysis of HA cases and contact persons in Erzinsky and Kyzyl regions of Tyva Republic was performed. Class M and G antibodies to HAV were determined in serum samples (n = 28), and HAV RNA--in stool samples (n = 16). Phylogenetic analysis of HAV RNA sequence was performed for VP1/2A region of the HAV genome with length 394 nucleotides. RESULTS: Cases of HA were registered during 3.5 months. Water supply sources did not have deviations from established standards. According to results of interviews, common food factor, which was able to cause the rise of HA incidence, was not determined. Signs of fecal contamination were revealed on environmental objects in preschool institutions and schools that demonstrate the low level of hygienic behaviour. It was shown that all cases of HA are related with different variants of the virus belonged to genotype IA that points to the absence of common source of infection. CONCLUSION: Results of epidemiological and genetic analysis of HAV demonstrate that observed rise in HA incidence in Tyva Republic are connected with phase of seasonal rise of HA incidence, which is characteristic for its perennial dynamics with active realization of contact route of virus transmission.