Národní referenční laboratoř pro stafylokoky CEM–SZÚ se i v roce 2019 v rámci zajištění surveillance stafylokokových infekcí věnovala podrobnému vyšetřování kmenů stafylokoků z humánního klinického materiálu. Celkem to bylo 1405 kmenů, převážně druhu Staphylococcus aureus, které byly zaslány asi z 80 bakteriologických pracovišť z celé České republiky. Metodou PCR byla zjišťována přítomnost genů kódujících především Pantonův-Valentinův leukocidin, toxin Syndromu toxického šoku, exfoliatiny a enterotoxiny. Informace o produkci faktorů virulence jsou důležité pro ošetřujících lékaře ke správnému stanovení diagnózy a tedy i vhodné terapie. V celém souboru bylo i 79 (5,6 %) kmenů koaguláza negativních stafylokoků. U těchto podmíněných patogenů jsme fenotypizací a metodou hmotnostní spektrometrie kmeny identifikovali, resp. konfirmovali identifikaci zjištěnou již v terénních laboratořích.

The main focus of the National Reference Laboratory for Staphylococci (NRL) in 2019 was again on the investigation of staphylococcal strains from human clinical specimens within the surveillance of staphylococcal infections. In total, 1405 strains mostly of the species Staphylococcus aureus referred to the NRL by 80 bacteriological laboratories from all over the Czech Republic were analysed. The strains were screened by PCR for the genes encoding Panton- -Valentine leucocidin, toxic shock syndrome toxin, exfoliatins, and enterotoxins. Data on the production of virulence factors are helpful for attending physicians in determining the right diagnosis and effective treatment. Seventy-nine strains (5.6%) referred to the NRL were coagulase negative staphylococci. These opportunistic pathogens were identified or confirmed, after previous identification by field laboratories, by phenotyping and mass spectrometry.

• MeSH
• Enterotoxins isolation & purification   classification   MeSH
• Exotoxins isolation & purification   MeSH
• Coagulase genetics   isolation & purification   MeSH
• Leukocidins isolation & purification   MeSH
• Humans MeSH
• Polymerase Chain Reaction methods   statistics & numerical data   MeSH
• Shock, Septic epidemiology   classification   microbiology   MeSH
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization methods   instrumentation   MeSH
• Staphylococcal Infections * epidemiology   genetics   classification   MeSH
• Staphylococcus genetics   isolation & purification   classification   pathogenicity   MeSH
• Check Tag
• Humans MeSH
• Geographicals
• Czech Republic MeSH

V programu surveillance byl v roce 2019 zjištěn v České republice mírný pokles počtu invazivních meningokokových onemocnění oproti předchozímu roku: celkem 49 případů (nemocnost 0,46/100 000 obyv.) oproti 56 v roce 2018 (nemocnost 0,52/100 000 obyv.) Z 49 onemocnění 3 skončila úmrtím – celková smrtnost v roce 2019 mírně stoupla ve srovnání s předchozím rokem na 6,1 % z 5,3 %. Úmrtí byla způsobena séroskupinami B a C – byla tedy preventabilní očkováním. Ve srovnání s předchozími roky pokračoval i v roce 2019 pokles procenta onemocnění způsobených N. meningitidis B (na 36,7 % ze 41,1 %), procento onemocnění způsobených N. meningitidis C zůstalo prakticky stejné v obou letech (42,9 % v roce 2019 a 42,8 % v roce předchozím). V roce 2019 zůstalo na podobných hodnotách jako v předchozím roce procento onemocnění způsobených séroskupinou W (6,1 % v roce 2019 a 7,1 % v roce předchozím) a séroskupinou Y (4,1 % v roce 2019 a 5,4 % v roce předchozím). U tří onemocnění nebyla v roce 2019 prokázána séroskupina: N. meningitidis ND (6,1 %). V roce 2019 kleslo procento invazivních meningokokových onemocnění prokázaných metodou PCR oproti předchozímu roku (na 30,6 % ze 35,7 %), u 24,5 % invazivních meningokokových onemocnění byla PCR jedinou metodou poskytující pozitivní výsledek. V roce 2019 byla v NRL provedena multilokusová sekvenční typizace (MLST) u všech kmenů z invazivního meningokokového onemocnění, které byly do NRL poslány. Nejčastěji zjištěným klonálním komplexem způsobujícím v roce 2019 invazivní onemocnění byl hypervirulentní komplex cc11, který patří mezi typické klonální komplexy séroskupiny C a W. Procento cc11 v roce 2019 ve srovnání s předchozím rokem mírně kleslo na 44,4 % ze 45,9 %.

The surveillance program data showed that the incidence of meningococcal invasive disease in the Czech Republic slightly decreased from 56 cases (0.52/100000) in 2018 to 49 cases (0.46/100000) in 2019. Three of the 49 cases were fatal, and the overall case fatality rate slightly increased from 5.3% in 2018 to 6.1% in 2019. The deaths were caused by N. meningitidis serogroups B and C and thus were vaccine preventable. In comparison to previous years, there was further decline in the proportion of cases caused by N. meningitidis B from 41.1% in 2018 to 36.7% in 2019 while the percentage of cases caused by N. meningitidis C remained almost unchanged, reaching 42.8% and 42.9%, respectively. Serogroup W cases remained similar in both years, with 7.1 % in 2018 and 6.1% in 2019 as well as serogroup Y cases, with 5.4% and 4.1% respectively. The causative serogroup was not determined in three cases (6.1%) in 2019 (N. meningitidis ND). The percentage of cases diagnosed by PCR decreased from 35.7 % in 2018 to 30.6 % in 2019. In 24.5% of cases, PCR was the only method to detect positivity. In 2019, the National Reference Laboratory for Meningococcal Infections performed multilocus sequence typing (MLST) of all referral strains from IMD. The most common causative hypervirulent complex involved in IMD in 2019 was cc11, typical for serogroups C and W. The proportion of cc11 cases showed a slight drop from 45.9% in 2018 to 44.9% in 2019.

• MeSH
• Humans MeSH
• Meningitis, Meningococcal * epidemiology   genetics   mortality   MeSH
• Meningococcal Vaccines therapeutic use   MeSH
• Morbidity MeSH
• Mortality MeSH
• Neisseria meningitidis, Serogroup B isolation & purification   MeSH
• Neisseria meningitidis, Serogroup C isolation & purification   MeSH
• Neisseria meningitidis, Serogroup W-135 isolation & purification   MeSH
• Polymerase Chain Reaction methods   statistics & numerical data   MeSH
• Age Distribution MeSH
• Check Tag
• Humans MeSH

Cíl: Ověření významu průkazu DNA Borrelia burgdorferi sensu lato u nervových forem lymeské boreliózy. Soubor a metodika: Bylo vyšetřeno 71 hospitalizovaných pacientů s prokázanou neuroboreliózou. Polymerázová řetězová reakce (PCR) byla vyšetřována paralelně v plazmě, moči a mozkomíšním moku před léčbou a po léčbě a dále po třech a šesti měsících v plazmě a moči. DNA byla izolována pomocí QIAamp DNA Mini Kitu. PCR probíhala jako dvoustupňová amplifikace (nested PCR). Každý biologický materiál byl testován paralelně pomocí pěti amplifikačních systémů: dva primery byly specifické pro plazmidové geny kódující OspA a OspC protein a tři pro chromozomální geny kódující 16S rDNA, flagelin a p66 protein. Výsledky: Z celkového počtu 71 pacientů byla alespoň v jedné tělní tekutině prokázána DNA před léčbou u 51 (71,8 %). Po léčbě byla DNA detekována u 25 pacientů (35,2 %). Po třech měsících pozitivita přetrvávala nadále u 19 (26,8 %) a po šesti měsících u pěti (7 %) nemocných. Nejvíce pozitivních výsledků bylo dosaženo pro 16S rDNA systém, o něco nižší citlivost byla zachycena s primery pro OspA, OspC a flagelin, nejnižší vykazoval p66 systém. Závěr: Boreliová DNA byla prokázána nejčastěji v mozkomíšním moku, méně v moči a nejmenší záchyt byl v plazmě. Pozitivita PCR během sledovaného období klesala postupně, u části pacientů byl test pozitivní ještě po šesti měsících. Z důvodu zlepšení diagnostické senzitivity lze doporučit paralelní vyšetření minimálně dvou cílových sekvencí. Na základě uvedených skutečností lze vyšetření DNA použít pouze jako doplňující test za předpokladu správné klinické interpretace jeho výsledků.

Aim: To assess the importance of Borrelia burgdorferi sensu lato DNA verification in neuroborreliosis. Material and methods: Seventy-one hospitalized patients with clinically and laboratory-confirmed neuroborreliosis were included. PCR of plasma, urine and CSF was performed in parallel before and after the treatment, and after 3 and 6 months for plasma and urine. DNA was isolated using the QIAamp DNA Mini Kit, PCR was arranged as a two-step amplification (nested PCR). Each sample was examined by the five amplification systems in parallel: two were targeted at plasmide genes encoding OspA and OspC proteins and three at genes for 16S rDNA, flagellin and p66 protein. Results: Of all 71 patients, the specific DNA was found in 51 cases (71.8%) before treatment. After the treatment, the DNA was proven in 25 patients (35.2%). PCR positivity persisted in 19 (26.8%) after three months and still in five (7%) patients after six months. The highest frequency of PCR-positive results was obtained by the 16S rDNA system, slightly lower sensitivity was achieved with the OspA, OspC and flagellin primers and the lowest frequency of positives was in the p66 target. Conclusion: Borrelial DNA was most frequently found in CSF, less frequently in urine and the lowest number of positive results were in plasma. PCR positivity decreased gradually during the observation period, the test was still positive in some patients after six months. It is possible to recommend testing with a minimum of two target sequences to improve diagnostic sensitivity. Our results suggest that the PCR testing should only be used in neuroborreliosis as an supplementary diagnostic method and subject to correct clinical interpretation.

• MeSH
• Borrelia burgdorferi * genetics   MeSH
• DNA, Bacterial genetics   blood   urine   MeSH
• DNA Primers * MeSH
• Humans MeSH
• Lyme Neuroborreliosis * genetics   blood   urine   MeSH
• Polymerase Chain Reaction statistics & numerical data   MeSH
• Prospective Studies MeSH
• Sensitivity and Specificity MeSH
• Check Tag
• Humans MeSH
• Publication type
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH

Faecal samples from 162 wild animals were collected from 32 distinct sites of Łęczyńsko-Włodawskie Lakeland (eastern Poland). The presence of Giardia duodenalis (Stiles, 1902) was assessed by a Direct Fluorescence Assay (DFA) and by Polymerase Chain Reaction (PCR) and sequencing of a fragment of the beta-giardin gene. DFA showed the presence of cysts of G. duodenalis in 12 of 162 faecal samples (7%), namely in four wild boars (15%), four foxes (19%), two roe deer (4%), and two wolves (29%). PCR identified 34 of the 162 (21%) samples as positive, including 11 wild boars (41%), five red deer (18%), 11 roe deer (23%), four moose (17%), two wolves (29%) and a single sample from the European badger. Thus, PCR detected a significantly higher number of infection than DFA (P = 0.0005). However, 14 of 34 PCR products could not be sequenced because of their insufficient amount; the low number of cysts, poor conservation of the faeces or presence of PCR inhibitors may have contributed to weak DNA amplification. Sequence analysis of the remaining 20 products showed the presence of assemblage B in wild boars, red deer and roe deer, whereas samples from wolves were identified as assemblage D. This is the first detection of assemblage B in wild boars and deer. As assemblage B has zoonotic potential, wild animals from eastern Poland may act as reservoirs of cysts of G. duodenalis infectious for humans.

• MeSH
• Animals, Wild classification   parasitology   MeSH
• Feces parasitology   MeSH
• Fluorescent Antibody Technique, Direct statistics & numerical data   MeSH
• Giardia lamblia * cytology   pathogenicity   MeSH
• Giardiasis epidemiology   etiology   MeSH
• Polymerase Chain Reaction statistics & numerical data   MeSH
• Sequence Analysis statistics & numerical data   MeSH
• Statistics as Topic MeSH
• Publication type
• Journal Article MeSH
• Geographicals
• Poland MeSH

Cílem práce bylo porovnání dosažených výsledků molekulárních metod využívaných v Oddělení bakteriálních vzdušných nákaz (OBVN) CEM SZÚ pro účely detekce, identifikace a typizace bakterií Neisseria meningitidis, Streptococcus pneumoniae a Haemophilus influenzae v klinických (kultivačně negativních) vzorcích při podezření na uvedená invazivní bakteriální onemocnění. OBVN nabízí využití molekulární diagnostiky uvedených bakteriálních původců menigitid a sepsí z klinických vzorků již řadu let. Spektrum metod se však změnilo. Z původní polymerázové řetězové reakce s elektroforetickým vyhodnocením (seminested PCR) k real-time polymerázové řetězové reakci (rt-PCR). Bylo provedeno souběžné srovnání obou metodik na 56 klinických vzorcích, které ukazuje lepší výsledky rt-PCR oproti seminested PCR.

The aim was to compare the outcomes of molecular methods used in the Unit for Airborne Bacterial Infections (UABI) for the detection, identification, and typing of the bacteria Neisseria meningitidis, Streptococcus pneumoniae, and Haemophilus influenzae from clinical culture-negative specimens of patients with suspected invasive bacterial infection. The UABI has been providing molecular diagnosis of the above-mentioned bacterial pathogens from clinical specimens of patients with meningitis or sepsis for years. However, the range of the methods used has changed: the UABI switched from the polymerase chain reaction (PCR) with reading of the results by elecrophoresis (seminested PCR) to the real-time polymerase chain reaction (rt-PCR). The outcomes of the two methods were compared using 56 clinical specimens, with rt-PCR proving superior to seminested PCR

• Keywords
• seminested PCR, rt-PCR,
• MeSH
• Molecular Diagnostic Techniques methods   statistics & numerical data   MeSH
• Haemophilus influenzae isolation & purification   MeSH
• Real-Time Polymerase Chain Reaction statistics & numerical data   MeSH
• Neisseria meningitidis isolation & purification   MeSH
• Polymerase Chain Reaction * methods   statistics & numerical data   MeSH
• Streptococcus pneumoniae isolation & purification   MeSH
• Bacterial Typing Techniques methods   statistics & numerical data   MeSH
• Publication type
• Comparative Study MeSH

The conventional polymerase chain reaction (PCR) method to detect the major allergenic protein parvalbumin beta 2 of Atlantic herring (Clupea harengus) and Pacific herring (Clupea pallasii) was developed. The specific set of primers for the amplification of the partial genomic sequence of the pvalb 2 gene encoding the main fish allergen of both herrings was designed and applied to the investigation of 24 commercial fish products. The targeted amplicon size was 189 bp of pvalb 2 gene of Atlantic herring and Pacific herring. As the internal amplification control, the DNA of 18S rRNA gene for eukaryotes (141 bp) was successfully used. The specificity of designed primer pair using 26 various fish species was assessed. The intrinsic detection limit was 10 pg µl(-1) of the present specific DNA. Atlantic herring or Pacific herring allergenic parvalbumins were detected in 22 investigated fish products in conformity with the package declaration. Two fish products were negative in spite of the declaration. The proposed PCR method is specific enough and can be used for the detection of Atlantic and Pacific herrings' major allergen parvalbumin beta 2 in fish food products.

• MeSH
• Allergens analysis   genetics   MeSH
• Food Contamination analysis   MeSH
• Humans MeSH
• Parvalbumins analysis   genetics   immunology   MeSH
• Polymerase Chain Reaction methods   statistics & numerical data   MeSH
• Food Hypersensitivity immunology   MeSH
• Fish Proteins analysis   genetics   immunology   MeSH
• Fish Products adverse effects   analysis   MeSH
• Fishes genetics   immunology   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Atlantic Ocean MeSH
• Pacific Ocean MeSH

Cíl práce: Cílem práce bylo zhodnotit epidemiologickou situaci kmenů komplexu Burkholderia cepacia izolovaných na Ústavu mi­krobiologie Lékařské fakulty Univerzity Palackého a Fakultní nemocnice Olomouc, zjistit nejčastější zástupce a prokázat, nebo vyvrátit možnost klonálního šíření těchto kmenů. Metodika: V průběhu osmi měsíců byly sbírány veškeré kmeny určené jako Burkholderia cepacia komplex. Byla stanovena citlivost na vybrané antimikrobiální přípravky a pomocí vhodných molekulárně genetických metod se zjišťovala jejich genetická příbuznost. Výsledky: Celkem bylo testováno 52 izolátů, nejčastěji (88,5 %) se jednalo o genomovar II (Burkholderia multivorans). Více než 46 % z nich bylo geneticky příbuzných a 58,3 % bylo zachyceno na jednotkách intenzivní péče. Všechny izoláty vykazovaly značnou rezistenci k antimikrobiálním přípravkům a úmrtí spojené s infekcí Burkholderia multivorans bylo popsáno ve čtyřech případech. Závěr: Lze předpokládat šíření geneticky příbuzných kmenů Burkholderia multivorans z nemocničního prostředí. Zdroj infekce zatím nebyl zjištěn a je třeba dalšího šetření.

Background: The aim was to assess the epidemiology of Burkholderia cepacia complex strains isolated at the Department of Microbiology, Faculty of Medicine and Dentistry, Palacký University and University Hospital Olomouc, determine the most frequent strains and confirm or rule out potential clonal spread of the strains. Material and methods: Over a period of eight months, all strains classified as Burkholderia cepacia complex were collected. Susceptibility to selected antimicrobial agents was determined and adequate molecular genetic methods were used to assess their genetic relationship. Results: A total of 52 isolates were tested, with the most frequent (88.5 %) being genomovar II (Burkholderia multivorans). More than 46 % of them were genetically related; 58.3 % of them were detected in intensive care units. All isolates were highly resistant to antimicrobial agents. In four cases, deaths associated with Burkholderia multivorans infection were reported. Conclusion: It may be assumed that genetically related strains of Burkholderia multivorans spread from the hospital setting. As yet, the source of infection has not been determined and further investigations are needed.

• Keywords
• Burkholderia multivorans, klonalita, rezistence,
• MeSH
• Drug Resistance, Bacterial genetics   MeSH
• Burkholderia cepacia complex genetics   isolation & purification   pathogenicity   MeSH
• Financing, Organized MeSH
• Genome, Bacterial genetics   MeSH
• Inpatients MeSH
• Burkholderia Infections epidemiology   microbiology   MeSH
• Cross Infection microbiology   MeSH
• Intensive Care Units MeSH
• Cohort Studies MeSH
• Humans MeSH
• Microbial Sensitivity Tests statistics & numerical data   MeSH
• Molecular Epidemiology methods   MeSH
• Polymerase Chain Reaction statistics & numerical data   MeSH
• Random Amplified Polymorphic DNA Technique statistics & numerical data   MeSH
• Check Tag
• Humans MeSH

BACKGROUND: Once the outbreak with Burkholderia cenocepacia ST32 was identified in the Prague cystic fibrosis (CF) centre, molecular tools were implemented into diagnostic routine in order to complement infection control measures with as accurate as possible microbiological service. In addition, genotyping techniques were applied as part of an infection surveillance program to assign species and strain status in samples positive for Burkholderia cepacia complex (Bcc). We sought to evaluate a usefulness of Bcc-specific PCR in infection control and to map evolution of the outbreak. METHODS: Since 2001, 6109 respiratory samples from 299 CF patients were examined not only by conventional culture, but also by PCR, detecting Bcc directly in sputum. RESULTS: Diagnosis of Bcc infection was established by culture in 54 patients already prior to 2001. As 39 more patients were diagnosed by culture and/or PCR during 2001-2010, this represented annual prevalence of 18.5%-28.9%. Twelve of 39 patients were culture negative at the time of their first PCR positivity. Although 2/3 of them became subsequently culture positive, the time delay in diagnostics of the infection by culture ranged from 1 to 22 months. New cases of Bcc infection were detected every year, however a dramatic drop was observed for the epidemic strain ST32. CONCLUSION: A likely factor contributing to the end of ST32 epidemic was segregation of Bcc infected patients that included also patients with no culture, but PCR positivity. The diagnostic PCR led to timely identification of cases with 'culture-invisible' infection.

• MeSH
• Burkholderia cepacia complex * genetics   isolation & purification   MeSH
• Cystic Fibrosis * epidemiology   microbiology   MeSH
• Adult MeSH
• Disease Outbreaks prevention & control   statistics & numerical data   MeSH
• Burkholderia Infections * diagnosis   epidemiology   prevention & control   MeSH
• Respiratory Tract Infections epidemiology   microbiology   prevention & control   MeSH
• Infection Control methods   MeSH
• Humans MeSH
• Polymerase Chain Reaction * methods   statistics & numerical data   MeSH
• Sputum microbiology   MeSH
• Bacterial Typing Techniques MeSH
• Check Tag
• Adult MeSH
• Humans MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Research Support, Non-U.S. Gov't MeSH
• Geographicals
• Czech Republic MeSH

Reliable and simple methods are required for detection of low concentrations of cytokines and some other proteins in complex biological fluids. This is especially important when monitoring the immune responses under various physiological and pathophysiological conditions in vivo or following production of these compounds in in vitro systems. Cytokines and other immunologically active molecules are being predominantly detected by enzyme-linked immunosorbent assays (ELISA) and newly also by immuno-polymerase chain reactions (iPCR). New simplified variants of iPCR have recently been described where antibodies are connected with multiple DNA templates through gold nanoparticles (Au-NPs) to form a new class of detection reagents. In this study we compared functionalized Au-NP-based iPCR (Nano-iPCR) with standard ELISA and iPCR for the detection of interleukin (IL)-3 and stem cell factor (SCF). The same immunoreagents (IL-3- and SCF-specific polyclonal antibodies and their biotinylated forms) were used throughout the assays. The obtained data indicate that both Nano-iPCR and iPCR are superior in sensitivity and detection range than ELISA. Furthermore, Nano-iPCR is easier to perform than the other two methods. Nano-iPCR was used for monitoring changes in concentration of free SCF during growth of mast cells in SCF-conditioned media. The results show that growing cultures gradually reduce the amount of SCF in supernatant to 25% after 5 days. The combined data indicate that Nano-iPCR assays may be preferable for rapid detection of low concentrations of cytokines in complex biological fluids.

• MeSH
• Cytokines analysis   immunology   MeSH
• Enzyme-Linked Immunosorbent Assay MeSH
• Immunoassay methods   statistics & numerical data   MeSH
• Indicators and Reagents MeSH
• Interleukin-3 analysis   immunology   MeSH
• Metal Nanoparticles MeSH
• Cells, Cultured MeSH
• Mast Cells immunology   MeSH
• Mice MeSH
• Nanotechnology MeSH
• Polymerase Chain Reaction methods   statistics & numerical data   MeSH
• Antibodies MeSH
• Stem Cell Factor analysis   immunology   MeSH
• Sensitivity and Specificity MeSH
• Gold MeSH
• Animals MeSH
• Check Tag
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH
• Evaluation Study MeSH
• Research Support, Non-U.S. Gov't MeSH
• Comparative Study MeSH

The study established the frequency of detection of antigens and antibodies to HBV and HCV in mixed-hepatitis B C. The highest rates of detection of serological markers were observed in the identification in the blood the HBV genotype D and HCV genotype 1b. This phenomenon can be explained by the high replicative and antigenic activity of viruses in these genotypes, resulting in a high viral load in blood, compared with HBV genotypes A and C, HCV genotypes 3a and 2a. In this regard, the use of ELISA technique in combination with PCR diagnosis and genotyping of viruses is considered as desirable for the etiological interpretation of viral hepatitis. This approach also will enhance achieving the correct diagnosis and adequate choice of treatment programs

• Keywords
• anti-HBs,
• MeSH
• Biomarkers blood   MeSH
• Hepatitis B, Chronic * genetics   immunology   MeSH
• Hepatitis C, Chronic * genetics   immunology   MeSH
• DNA, Viral MeSH
• Enzyme-Linked Immunosorbent Assay statistics & numerical data   MeSH
• Genotype MeSH
• Hepacivirus genetics   immunology   MeSH
• Hepatitis B e Antigens genetics   immunology   MeSH
• Hepatitis B Core Antigens genetics   immunology   MeSH
• Hepatitis B Surface Antigens genetics   immunology   MeSH
• Hepatitis C Antigens genetics   immunology   MeSH
• Immunoglobulin G analysis   immunology   MeSH
• Immunoglobulin M analysis   immunology   MeSH
• Liver Cirrhosis pathology   MeSH
• Biopsy, Needle MeSH
• Coinfection * genetics   immunology   MeSH
• Humans MeSH
• Polymerase Chain Reaction statistics & numerical data   MeSH
• RNA, Viral MeSH
• Viral Load statistics & numerical data   MeSH
• Genes, Viral * MeSH
• Hepatitis B virus genetics   immunology   MeSH
• Check Tag
• Humans MeSH
• Geographicals
• Uzbekistan MeSH