BACKGROUND: Single nucleotide polymorphisms can create a genetic microenvironment in some tumors that affects the course of treatment, resistance, etc. Whether single nucleotide polymorphisms have an impact on gastrointestinal stromal tumor (GIST) development and disease progression is not yet accurately verified. KIT SNPM541L in exon 10 correlates with a worse prognosis of many cancers. The impact of KIT SNPM541L in GISTs is relatively unknown and, therefore, its analyses could have potential in patient therapy and could provide more detailed information on tumor character, clinical presentation, or tumor behavior in treatment. AIM: The aim of the study was the analysis of the biological and clinical significance of the KIT SNPM541L polymorphism in exon 10. MATERIALS AND METHODS: Paraffin sample tissues were obtained from the National GIST Register in Martin. Retrospective samples from 177 GIST patients were divided into several groups. Detection of SNPM541L was performed by Sanger sequencing. Statisitical analyses were performed to determine the prevalence of KIT SNPM541L in the Slovak GIST cohort, to search for correlation between c-KIT status and clinicopathological, molecular and biological data. RESULTS: Overall, 29 samples out of 177 showed KIT SNPM541L polymorphism. CONCLUSION: Our results do not support the association between KIT SNPM541L and increased risk of relapse in localized primary GISTs. Additionally, we found a positive correlation between KIT SNPM541L occurrence and earlier onset of relapse in PDGFRa and WT subgroup of GISTs.
- MeSH
- dospělí MeSH
- gastrointestinální nádory epidemiologie genetika patologie MeSH
- gastrointestinální stromální tumory epidemiologie genetika patologie MeSH
- jednonukleotidový polymorfismus * MeSH
- lidé středního věku MeSH
- lidé MeSH
- míra přežití MeSH
- mladiství MeSH
- mladý dospělý MeSH
- nádorové biomarkery genetika MeSH
- následné studie MeSH
- prognóza MeSH
- protoonkogenní proteiny c-kit genetika MeSH
- retrospektivní studie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Evropa MeSH
Východiská: Podstatou moderných postupov liečby onkologických pacientov je v dnešnej dobe zacielenie konkrétnych molekúl zapojených do bunkovej signalizácie asociovanej s nádorovou iniciáciou a progresiou. Úspech uvedeného prístupu závisí od správne zvoleného diagnostického testu s vysokou citlivosťou, ktorý identifikuje výskyt a hladinu vybraných biomarkerov u pacientov pre selekciu tých, ktorí budú na liečivo reagovať a budú z neho benefitovať. Vývoj nových technológií a modernizácia tých známych, prispievajú k inováciám molekulárnej charakterizácie karcinómov, ktorá umožňuje detekciu mutačného stavu pacienta s vysokou citlivosťou a špecifickosťou. Cieľ: V práci diskutujeme o využití polymerázovej reťazovej reakcie (PCR) tretej generácie, tzv. droplet digitálnej PCR (ddPCR), v molekulárnej diagnostike karcinómov. Podľa štúdií uvedených v našom prehľade predstavuje ddPCR sľubný nástroj pri vytváraní genetického profilu pacientov s onkologickým ochorením. Optimalizácia a presná validácia môžu preto umožniť postupnú implementáciu ddPCR do klinickej praxe v oblasti onkológie.
Background: Nowadays, modern treatment methods for cancer patients are based on targeting specific molecules involved in cellular signaling system associated with tumor initiation and progression. The success of such approach depends on a correctly chosen diagnostic test with high sensitivity that identifies the occurrence and level of biomarkers in patients to select those who will respond and benefit from the treatment. The development of new technologies and the upgrades of the known ones contribute to the innovations in molecular characterization of cancer, which allows the detection of patient’s mutational status with high sensitivity and specificity. Purpose: Here, we discuss the utilization of the third-generation type of polymerase chain reaction (PCR), droplet digital PCR (ddPCR), in the molecular diagnostics of oncology diseases. According to the studies reported in our review, ddPCR represents a promising tool in genetic profiling of cancer patients. Therefore, the optimization and precise validation may enable gradual implementation of ddPCR into clinical practice in the field of oncology.
The proto-oncogene KRAS belongs among the most frequently mutated genes in all types of cancer and is also very important oncogene related to colorectal tumors. The detection of mutations in this gene in primary tumor is a predictive biomarker for the anti-EGFR therapy in metastatic CRC (mCRC); however, the patients with wild-type KRAS can also show resistance to the personalized medicine. The droplet-based digital PCR technology has improved the analytical sensitivity of the mutations detection, which led us to the idea about the optimization of this approach for KRAS testing. In this study, we report the application of ddPCR technology in order to analyze the presence of KRAS mutations in primary tumor and matched metastasis in lymph nodes (LNs) from patients with mCRC and address the question, whether the improvement in the detection method can lower the discrepancies of KRAS mutations detection between the primary tumor and regional LNs. Genomic DNA with wtKRAS and commercial DNA with mtKRAS (G12D) were used to set up the ddPCR reaction. Formalin-fixed paraffin-embedded tissues from primary tumor and positive lymph node from 31 patients with mCRC were analyzed using ddPCR and Sanger sequencing. KRAS status of primary tumors was known; however, the mutation status of lymph nodes was not detected previously. From 31 samples of primary tumors, our results corresponded to results from IVD kit in 30 cases. For one patient, ddPCR detected KRAS mutation in comparison with negative result of the IVD kit. In the samples of metastatic infiltrated LNs, ddPCR detected 16 samples as a WT KRAS and 15 lymph nodes showed positivity for KRAS mutation, whereby Sanger sequencing found KRAS mutations in 8 cases only. We also found two cases where genetic conditions of KRAS gene differed between primary tumor and infiltrated lymph node, both "low-grade" adenocarcinoma. Our study approved that ddPCR method is adequate technique with high sensitivity and in the future may be used as a diagnostic tool for evaluation of KRAS mutations, especially in infiltrated LNs of patients with mCRC.
- MeSH
- kolon patologie MeSH
- kolorektální nádory diagnóza genetika patologie MeSH
- lidé MeSH
- lymfatické uzliny patologie MeSH
- molekulární patologie metody MeSH
- mutantní proteiny genetika MeSH
- polymerázová řetězová reakce metody MeSH
- protoonkogenní proteiny p21(ras) genetika MeSH
- senioři MeSH
- senzitivita a specificita MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
Tibetans existed in high altitude for ~25 thousand years and have evolutionary selected unique haplotypes assumed to be beneficial to hypoxic adaptation. EGLN1/PHD2 and EPAS1/HIF-2α, both crucial components of hypoxia sensing, are the two best-established loci contributing to high altitude adaptation. The co-adapted Tibetan-specific haplotype encoding for PHD2:p.[D4E/C127S] promotes increased HIF degradation under hypoxic conditions. The Tibetan-specific 200 kb EPAS1 haplotype introgressed from an archaic human population related to Denisovans which underwent evolutionary decay; however, the functional variant(s) responsible for high-altitude adaptation at EPAS1/HIF-2α have not yet been identified. Since HIF modulates the behavior of cancer cells, we hypothesized that these Tibetan selected genomic variants may modify cancer risk predisposition. Here, we ascertained the frequencies of EGLN1D4E/C127S and EGLN1C127S variants and ten EPAS1/HIF-2α variants in lung cancer patients and controls in Nepal, whose population consists of people with Indo-Aryan origin and Tibetan-related Mongoloid origin. We observed a significant association between the selected Tibetan EGLN1/PHD2 haplotype and lung cancer (p=0.0012 for D4E, p=0.0002 for C127S), corresponding to a two-fold increase in lung cancer risk. We also observed a two-fold or greater increased risk for two of the ten EPAS1/HIF-2α variants, although the association was not significant after correcting for multiple comparisons (p=0.12). Although these data cannot address the role of these genetic variants on lung cancer initiation or progression, we conclude that some selected Tibetan variants are strongly associated with a modified risk of lung cancer.
- MeSH
- aklimatizace MeSH
- faktor 1 indukovatelný hypoxií - podjednotka alfa genetika metabolismus MeSH
- hypoxie genetika metabolismus MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádory plic genetika metabolismus MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Tibet MeSH
Tibetans have lived at high altitude for generations and are thought to be genetically adapted to hypoxic environments. Most are protected from hypoxia-induced polycythemia, and a haplotype of EPAS1, encoding hypoxia-inducible factor (HIF-2α), has been associated with lower hemoglobin levels. We earlier reported a Tibetan-specific EGLN1 haplotype encoding PHD2 which abrogates HIF augmentation in hypoxia. We genotyped 347 Tibetan individuals from varying altitudes for both the Tibetan-specific EGLN1 haplotype and 10 candidate SNPs in the EPAS1 haplotype and correlated their association with hemoglobin levels. The effect of the EGLN1 haplotype on hemoglobin exhibited age dependency at low altitude, while at higher altitudes, it showed a trend to lower hemoglobin levels in the presence of the Tibetan-selected EPAS1 rs142764723 C/C allele. The observed gene-environment and gene-gene interactions and the moderate effect of the EGLN1 and EPAS1 haplotypes on hemoglobin indicate that other modifiers exist. It remains to be determined whether a blunting of erythropoiesis or other physiological consequences of HIF downregulation are the primary drivers of these genetic adaptations among Tibetans. KEY MESSAGE: Most Tibetans are protected from polycythemia while living in high altitude. An EGLN1 co-adapted haplotype, EGLN1 c.12C>G, c.380G>C is uniquely Tibetan. The Tibetan EPAS1 haplotype has introgressed from the Denisovan genome. While EGLN1 and EPAS1 genotypes lower Hb, this study indicates additional Hb modifiers.
- MeSH
- aklimatizace genetika MeSH
- Asijci genetika MeSH
- dospělí MeSH
- erythropoetin krev MeSH
- ferritin krev MeSH
- haplotypy MeSH
- hemoglobiny analýza MeSH
- interakce genů a prostředí MeSH
- jednonukleotidový polymorfismus MeSH
- lidé MeSH
- nadmořská výška MeSH
- prolyl-4-hydroxylasy HIF genetika MeSH
- transkripční faktory bHLH genetika MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Tibet MeSH
INTRODUCTION: Episodic ataxias (EAs) are rare dominantly inherited neurological disorders characterized by recurrent episodes of ataxia lasting minutes to hours. The most common subtype is EA type 2 (EA2) caused by pathogenic variants of calcium voltage-gated channel subunit alpha1 A gene (CACNA1A) on chromosome 19p13. SUBJECTS AND METHODS: We examined a Slovak three-generation family. Genomic DNA of the family members was extracted from peripheral blood and amplified by polymerase chain reaction. CACNA1A variants were screened by Sanger sequencing. RESULTS: We identified four family members with recurrent episodes of ataxia. Complex differential diagnosis was performed. Genetic analysis with direct sequencing revealed a novel heterozygous variant of CACNA1A - c.5264A>G (p.Glu1755Gly) located in the pore loop of domain IV of calcium channel alpha-1A subunit. CONCLUSION: We identified a novel missense variant of a voltage-dependent P/Q-type calcium channel alpha-1A subunit in a Slovak three-generation family with recurrent episodes of ataxia. The heterozygous missense variant resulted in changing a highly conserved glutamic acid within the pore loop of domain IV.
- MeSH
- ataxie genetika MeSH
- heterozygot MeSH
- lidé středního věku MeSH
- lidé MeSH
- missense mutace genetika MeSH
- mladý dospělý MeSH
- recidiva MeSH
- rodokmen MeSH
- vápníkové kanály genetika MeSH
- věk při počátku nemoci MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
- Geografické názvy
- Slovenská republika MeSH
Cieľ štúdie: Zavedenie metodiky QF-PCR pre detekciu najčastejšie sa vyskytujúcich chromozómových (trizómia 21, 18 a 13) a gonozómových aberácií na našom pracovisku z plodovej vody získanej pri amniocentéze v druhom trimestri. Overenie hypotézy možnosti detekcie chromozómových aberácií použitím STR markerov kitu Aneufast analýzou voľnej fetálnej DNA (ffDNA) izolovanej z plazmy matky s plodom postihnutým trizómiou 21. Typ štúdie: Prospektívna klinická štúdia. Názov a sídlo pracoviska: Gynekologicko-pôrodnícka klinika Jesseniovej lekárskej fakulty Univerzity Komenského a Univerzitnej Nemocnice Martin, Slovenská republika. Metodika: Vzorky plodovej boli získané od 67 žien (3 prípady dvojplodovej tehotnosti) v druhom trimestri gravidity (15. až 22. týždeň tehotnosti (t.t.)) boli vyšetrené multiplexnou QF-PCR kitom Aneufast a v prípade pozitivity pre trizómiu 21 aj kitom Devyser Resolution 21. Všetky vzorky boli zároveň vyšetrené konvenčnou cytogenetickou karyotypizáciou. Analýza ffDNA kitom Aneufast z plazmy 3 vysokorizikových žien odobratej v druhom trimestri (17. až 21. t.t.) prebehla po izolácii ffDNA kitom QIAamp DSP VirusKit. Trizómia 21 bola u týchto plodov potvrdená karyotypizáciou amniocytov v druhom trimestri. Výsledky: V súbore 70 vzoriek vyšetrenom pomocou QF-PCR bolo celkovo zachytených 7 patologických vzoriek z toho 6 trizómií 21 a 1 trizómia 18. U všetkých vzoriek vyšetrených metódou QF-PCR došlo ku 100% zhode s výsledkami cytogenetickej karyotypizácie. V prípade analýzy ffDNA nedošlo ku dostatočnej a teda interpretovateľnej amplifikácii potrebných úsekov chromozómu 21. Záver: QF-PCR sa ukázala ako spoľahlivá, rýchla, pomerne jednoduchá a finančne prijateľná metodika prenatálnej diagnostiky. Napriek dostupnosti a dobrému zavedeniu práce s kitom Aneufast, výsledky analýzy ffDNA nie sú uspokojivé. Vyšetrením ffDNA izolovanej z plazmy 3 žien s plodmi s trizómiou 21 sme nezískali interpretovateľné výsledky.
Objective: To introduce QF-PCR method for detection of the most common chromosomal (trisomy 21, 18 and 13) and gonosomal aneuploidies at our department in the second–trimester amniotic fluid. To test the hypothesis of chromosomal aneuploidies detection using STR markers of Aneufast? kit via analysing free fetal DNA (ffDNA) isolated from plasma of pregnant women with confirmed trisomy 21 in fetus. Design: A prospective clinical study. Setting: Department of Obstetrics and Gynecology, Jessenius Faculty of Medicine and University Hospital in Martin, Slovak Republic. Methods: The samples of amniotic fluid were obtained from 67 women (twin pregnancy in 3 cases) in the 2nd trimester (15th to 22nd gestational week (g.w.)). Samples were examined using multiplex QF-PCR via Aneufast kit. In the case of positivity for trisomy 21, they were re-examined using Devyser Resolution 21 kit. All samples were parallelly evaluated by cytogenetic karyotyping. We also analyzed ffDNA from the plasma of 3 high-risk women using Aneufast kit. The plasma samples were obtained in the 2nd trimester(17th to 21st g.w.). Qiaamp DSP Virus kit was used for ffDNA isolation. Trisomy 21 of 3 fetuses was confirmed by karyotyping after 2nd trimester amniocentesis. Results: In the cohort of 70 samples, 7 pathological results (six trisomies 21 and one trisomy 18) were obtained. There was 100% concordance with cytogenetic karyotype in all samples examined by QF-PCR. The amplification of tracked chromosome 21 fragments was not evaluable in the case of ffDNA analysis. Conclusion: QF-PCR was approved as reliable, rapid, quite simple and financially bearable method of prenatal diagnostics. Despite the fact of good availability and work implementation of Aneufast? kit, results of ffDNA analysis are insufficient. We did not obtain interpretable results after ffDNA analysis from maternal plasma in trisomy 21 fetuses.
- Klíčová slova
- QF-PCR, ffDNA,
- MeSH
- chromozomální aberace MeSH
- cytogenetika MeSH
- DNA * analýza MeSH
- dospělí MeSH
- druhý trimestr těhotenství MeSH
- karyotypizace MeSH
- krevní plazma MeSH
- lidé MeSH
- plod * MeSH
- polymerázová řetězová reakce * metody využití MeSH
- prenatální diagnóza MeSH
- trizomie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
Klasifikácia SZO (2008) je jediná celosvetovo akceptovaná a v praxi používaná klasifikácia nádorových ochorení kostnej drene (KD), ktorá definuje aj postavenie a význam biopsie KD v diagnostickom algoritme. Myeloproliferatívne neoplázie (MPN) tvoria pestrú skupinu ochorení s variabilne dynamickým rozvojom z počiatočných štádií, cez štádiá s plno rozvinutou klinicko-laboratórnou manifestáciou až do pokročilých štádií – fibrotickej, resp. blastovej transformácie, sprevádzaných zmenami morfológie a laboratórneho obrazu. V počiatočných štádiách je určenie typu MPN len na základe klinicko-laboratórnych údajov pre podobnosť ich klinickej manifestácie nespoľahlivé. Dôkaz pre MPN typických genetických mutácií potvrdzuje klonalitu ochorenia, ale jeho typizačný význam je limitovaný. Naopak, bioptické vyšetrenie KD v kontexte dostatočných klinických informácií, výsledkov laboratórnych vyšetrení a s podporou genetických vyšetrení umožňuje okrem potvrdenia MPN aj bližšiu typizáciu, ktorá je prognosticky relevantná a ovplyvňuje aj voľbu terapie. Zároveň umožňuje odlíšenie reaktívnych zmien hemopoézy, zhodnotenie prognosticky významných morfologických znakov (stupeň myelofibrózy, množstvo blastov) a sledovanie progresie ochorenia do fibrotickej, resp. blastovej transformácie. V týchto pokročilých štádiách a po terapii nie je však typizácia MPN len na základe morfológie KD možná. Výpovedná hodnota biopsie závisí od kvality a kvantity vzorky drene, hodnotenie morfológie KD závisí aj od vedomostí a skúseností vyšetrujúceho patológa.
WHO classification (2008) is the only world-wide accepted and in routine praxis used classification of bone marrow (BM) neoplasms defining also the importance of BM biopsy within the diagnostic algorithm. Myeloproliferative neoplasias (MPN) represent a heterogenous group of disorders with variable dynamic transformational potential from initial into advanced stages such as blast crisis or myelofibrotic transformation. Because of similar manifestations, the typing of MPN in their initial stages based on clinical and laboratory parameters is not optimal. The finding of MPN-typical genetic mutations proves the disease clonality, but not a diagnosis of individual MPN types. In contrast, the BM biopsy verifies both the diagnosis and MPN type. It helps also to discriminate MPN from reactive hematopoietic changes, to evaluate prognostically relevant BM features (e.g. grade of myelofibrosis, blasts number) and the follow-up to terminal stages. However, the MPN typing in terminal and posttherapeutical stages based on morphology alone is not sufficient. The diagnostic value of BM biopsy depends on specimen´s quality, the objectivity of BM morphology evaluation depends on pathologist´s training and experiences. A correct BM biopsy interpretation should be done within the context of sufficient clinical and laboratory data and with support of genetical analyses. myelofibrosis, bone marrow biopsy.
- MeSH
- biopsie využití MeSH
- diferenciální diagnóza MeSH
- esenciální trombocytemie diagnóza patologie MeSH
- financování organizované MeSH
- hematologie MeSH
- histologické techniky využití MeSH
- kostní dřeň patologie MeSH
- leukemie myeloidní chronická atypická BCR-ABL-negativní diagnóza patologie MeSH
- lidé MeSH
- megakaryocyty patologie MeSH
- myeloproliferativní poruchy diagnóza patologie MeSH
- polycythaemia vera diagnóza patologie MeSH
- primární myelofibróza diagnóza patologie MeSH
- vyšetřování kostní dřeně MeSH
- Check Tag
- lidé MeSH
Polycythemia vera (PV), esenciálna trombocytóza (ET) a primárna myelofibróza (PMF) patria do skupiny Ph1 negatívnych myeloproliferatívnych neoplázii (MPN), pri ktorých sa často vyskytuje mutácia JAK2V617F. Táto mutácia sa vyskytuje u takmer všetkých pacientov s PV a u približne 60 % pacientov s ET a PMF. V diferenciálnej diagnostike MPN však význam tejto mutácie stále nie je dosť objasnený, preto tu má dôležitú úlohu aj bioptické vyšetrenie kostnej drene. Na našom pracovisku sme sa rozhodli vyšetrovať JAK2V617F mutáciu v DNA izolovanej z parafínovych blokov od pacientov s PV, ET a PMF, keďže túto mutáciu nedávno SZO odporučila využívať ako jeden z markerov v diagnostike uvedených klinických jednotiek. Mutácie V617F sme detekovali pomocou alelovo-špecifickej hot start multiplex PCR. U JAK2V617F negatívnych pacientov s PV sme vyšetrovali sekvenovaním prítomnosť mutácií v exóne 12 JAK2 génu. Dosiaľ sme na našom pracovisku vyšetrili približne 200 pacientov s klinicky verifikovanou diagnózou PV, ET a PMF. Naše výsledky sú u všetkých troch ochorení (ET, PV, PMF) zhodné s dosiaľ publikovanými prácami. Nami implementovaná metodika detekcie mutácie JAK2 z parafínových blokov umožňuje analýzu veľkého množstva archívneho materiálu pre retrospektívne štúdie, a súčasne aj implementáciu analýzy statusu JAK2 ako súčasti bioptického vyšetrenia kostnej drene všetkých pacientov s podozrením na PV, ET alebo PMF.
Polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF) are Philadelphia chromosome-negative myeloproliferative neoplasms (MPN) characterized by JAK2 mutation. The exon 14 V617F mutation is present in almost all patients with PV and in approx. 60¨% of patients with ET and PMF. The importance of JAK2V617F in the differential diagnostic considerations is still unclear and here the BM morphology examination still represents an important diagnostic tool. In the WHO classification of Ph1-negative MPNs, the identification of JAK2 mutations represents a major diagnostic criterion of these diseases. Therefore we decided to implement the examination of JAK2V617F mutation in formalin-fixed paraffin-embedded biopsy specimens of patients with Ph1-negative MPN using allele-specific PCR. In addition, in all JAK2 V617F negative patients with PV we sequenced the whole JAK2 exon 12. Until now we examined up to 200 patients with clinically confirmed MPN and our results in all three categories PV, ET and PMF are in agreement with earlier published data. Paraffin embedded tissues represent a valuable source of DNA which can be used in the diagnostics of both JAK2 exon 12 and exon 14 mutations. It is of particular importance if the fresh material is not available and there is a clinical and/or research utility for the performance of PCR on archival bone marrow samples with PV, ET or PMF suspicion.
- MeSH
- biopsie metody využití MeSH
- bodová mutace genetika imunologie MeSH
- diagnostické techniky molekulární využití MeSH
- DNA genetika izolace a purifikace MeSH
- esenciální trombocytemie diagnóza genetika MeSH
- financování organizované MeSH
- Janus kinasa 2 genetika izolace a purifikace MeSH
- lidé MeSH
- myeloproliferativní poruchy diagnóza etiologie genetika MeSH
- polycythaemia vera diagnóza etiologie genetika MeSH
- polymerázová řetězová reakce metody využití MeSH
- primární myelofibróza diagnóza genetika MeSH
- retrospektivní studie MeSH
- vyšetřování kostní dřeně metody využití MeSH
- Check Tag
- lidé MeSH
