Helicobacter pylori is one of the most common cause of human infections. Infected patients develop chronic active gastritis in all cases, which can lead to peptic ulcer, atrophic gastritis, gastric cancer and gastric MALT-lymphoma. The prevalence of H. pylori infection in the population has regional characteristics and can reach 80%. Constantly increasing antibiotic resistance of H. pylori is a major cause of treatment failure and a major problem. According to the VI Maastricht Consensus, two main strategies for choosing eradication therapy are recommended: individualized based on evaluating sensitivity to antibacterial drugs (phenotypic or molecular genetic method) prior to their appointment, and empirical, which takes into account data on local H. pylori resistance to clarithromycin and monitoring effectiveness schemes in the region. Therefore, the determination of H. pylori resistance to antibiotics, especially clarithromycin, prior to choosing therapeutic strategy is extremely important for the implementation of these treatment regimens.

The Maastricht VI/Florence consensus recommends, as one of the measures to enhance the efficacy of Helicobacter pylori infection eradication, a personalized treatment approach involving the selection of an antimicrobial agent based on the pre-determined resistance of H. pylori. To address the need to develop test systems for personalized drug selection, this study was designed to analyze the molecular resistance of H. pylori using a newly developed Sanger sequencing test platform. The characteristics of the test system were determined on 25 pure culture samples of H. pylori with known resistance. Sensitivity and specificity for detecting resistance to clarithromycin was 100% and those to levofloxacin were 93% and 92%, respectively. The test system has been tested in real clinical practice on 112 H. pylori-positive patients who had not previously received proton pump inhibitors (PPIs) or antibacterial drugs. Mutations indicating resistance to clarithromycin were found in 27 (24%) samples and those indicating resistance to levofloxacin were found in 26 (23%) samples. Double resistance was observed in 16 (14%) samples. The most common mutations leading to clarithromycin resistance were 2143G and 2142G and to levofloxacin resistance—261A and 271A in the gyrA gene, which account for 69% of all identified genetic determinants in levofloxacin-resistant bacteria. Thus, a personalized approach to the selection of H. pylori eradication therapy based on the detection of bacterial resistance before prescribing first-line therapy could help to avoid the prescription of ineffective H. pylori eradication therapies and, overall, contribute to the control of antibiotic resistance of H. pylori.

Aim. To evaluate the possibility of using serum markers of atrophy (pepsinogens – PG I and II) to form high-risk groups for gastric cancer (Operative Link for Gastritis Assessment – OLGA stage III–IV) depending on the etiology of gastritis. Materials and methods. A total of 237 (56 men and 181 women) patients were examined. All patients underwent a 13C-urea breath test, a blood test for GastroPanel (PG I, PG II, gastrin-17, antibodies to Helicobacter pylori immunoglobulin G), a blood test for antibodies to gastric parietal cells. All patients underwent esophagogastroduodenoscopy with a biopsy of the gastric mucosa from 5 standard points according to the Sydney system and a histomorphological study according to the OLGA system, as well as a biopsy to detect H. pylori infection using the polymerase chain reaction. The patients were divided into 3 groups depending on the etiology of gastritis: Group 1 included 55 patients with chronic gastritis, autoimmune gastritis and associated with H. pylori gastritis (AIG+HP+); Group 2 – 47 patients with AIG and negative tests for H. pylori infection (AIG+HP-); Group 3 – 135 patients with chronic gastritis associated with H. pylori and negative markers of AIG (AIG-HP+). Results. The analysis showed that in patients with AIG (group 2), the most reliable serological markers of atrophy predicted severe atrophy (OLGA stage III–IV): when the ratio PG I/PG II was ≤ 3, it was detected in 70.21% of cases, and when PG I decreased to ≤ 30 μg/L, it was found in 68.08%. In group 1, stages III–IV according to OLGA were diagnosed in 20% of cases with PG I/PG II indicators ≤ 3; and in 18.18% with a decrease in PG I ≤ 30 μg/L. When analyzing the diagnostic accuracy of GastroPanel biomarkers in identifying severe atrophy (OLGA stages III–IV) in the total sample of patients (all 3 groups), it was possible to achieve cut-off indicators as close as possible to the reference values while maintaining a relatively high sensitivity and specificity – 75.81% and 81.50% for PG I ≤ 30 μg/L and 85.48% and 64.50% for PG I/PG II ≤ 3, respectively. The optimal cut-off in the study population for the PG I indicator was 22.5 μg/L (sensitivity – 72.58%, specificity – 88.00%), and for the PG I/PG II ratio ≤ 2 (sensitivity – 80.65%, specificity – 78.50%). Conclusion. Serum pepsinogens can be used in the Moscow population as a non-invasive marker of gastric mucosa atrophy for the formation of high-risk patient groups for gastric cancer requiring endoscopic examination.

В результате формирования устойчивости H. pylori к антибиотикам снизилась эффективность эрадикационной терапии. Выбор схем эрадикации базируется на данных о локальной резистентности к антибиотикам, прежде всего к кларитромицину. Предложен индивидуализированный подход эрадикации H. pylori, на основе определения устойчивости к антибиотикам до назначения терапии первой линии. ЦЕЛЬ ИССЛЕДОВАНИЯ Провести сравнительную характеристику фенотипического и молекулярно-генетических методов выявления антибиотикорезистентности H. pylori, оценить их диагностическую чувствительность и специфичность. МАТЕРИАЛ И МЕТОДЫ В исследование были включены 25 чистых культур H. pylori с установленной фенотипической чувствительностью и устойчивостью к антибактериальным препаратам. Устойчивость к кларитромицину и левофлоксацину определяли путем серийных разведений в катион-сбалансированном агаре Мюллера—Хинтон с добавлением овечьей крови в соответствии с рекомендациями Института клинических и лабораторных стандартов США (CLSI). Молекулярные маркеры устойчивости оценивали с помощью секвенирования по Сэнгеру и полимеразной цепной реакции в реальном времени (RT-PCR) со специфичными праймерами. Диагностическую чувствительность и специфичность молекулярных методов определяли в сравнении с результатами фенотипического метода. РЕЗУЛЬТАТЫ Установлено полное соответствие результатов фенотипических и молекулярных методов выявления устойчивости H. pylori к кларитромицину. При оценке резистентности к левофлоксацину было показано, что культуры, демонстрирующие фенотипическую устойчивость, также содержали мутации в гене gyrA, связанном с устойчивостью. Вместе с тем, некоторые культуры без фенотипической устойчивости имели характерные мутации гена gyrA. Чувствительность и специфичность молекулярных методов определения резистентности к кларитромицину составила 100%. Чувствительность выявления устойчивости к левофлоксацину составила 100%, специфичность — 78,57%. ВЫВОДЫ Исследование демонстрирует сильную связь между фенотипической и молекулярной резистентностью H. pylori, особенно к кларитромицину. Секвенирование по Сэнгеру и RT-PCR дали стабильные результаты, что подчеркивает надежность данного метода.

This paper documents a case report of a rare aggressive variant of Cyclin D1-negative mantle cell lymphoma combined with a congenital anomaly, one of the 46,XY disorders of sex development (DSD). In an elderly female patient aged 71 years, histological and immunohistochemical analyses of the bone marrow revealed a diffuse infiltration of atypical CD5-positive Cyclin D1-negative lymphoid cells with blastoid morphology expressing SOX11 as well as high proliferative index Ki-67 ≤ 70 %. In the cytogenomic profile of the tumor, the translocation t(11;14)(q13;q32) CCND1::IGH was not identified, however, it was marked by multiple clonal chromosome abnormalities including the deletion of the 17p13/TP53 locus and the involvement of the 14q32/IGH locus in the translocation with an unidentified partner. At the same time, the ХY(SRY+) gonosomal complement was detected in the patient’s bone marrow cells, peripheral blood lymphocytes, and buccal epithelial cells, enabling, for the first time, to diagnose a congenital anomaly classified as a 46,XY DSD. Despite chemotherapy (4 VR-CAP cycles and Bruton tyrosine kinase inhibitors), rapid progression of the underlying disease led to the patient’s death. The paper reports diagnostic and clinical features of Cyclin D1-negative mantle cell lymphoma and discusses a possible relationship between DSD and some hematologic tumors.

The Maastricht VI/Florence consensus recommends, as one of the measures to enhance the efficacy of Helicobacter pylori (H. pylori) infection eradication, a personalized treatment approach involving the selection of an antimicrobial agent based on the pre-determined resistance of H. pylori. To meet the need to develop test systems for personalized drug selection, we conducted a study of H. pylori molecular resistance in Moscow using a self-developed Sanger sequencing test platform. We validated our test system on 25 pure culture samples of H. pylori with known resistance. Sensitivity and specificity for detecting resistance to clarithromycin was 100%, and to levofloxacin - 93% and 92%, respectively. Then we tested our platform on 112 H. pylori -positive patients who had not previously received proton pump inhibitors (PPI) or antibacterial drugs. Mutations indicating resistance to clarithromycin were found in 27 (24%) samples, and to levofloxacin in 26 (23%) samples. Double resistance was observed in 16 (14%) samples. The most common mutations leading to clarithromycin resistance were 2143G and 2142G, to levofloxacin resistance – 261A and 271A in the gyrA gene, which account for 69% of all identified genetic determinants in levofloxacin-resistant bacteria. Thus, a personalized approach to the selection of H. pylori eradication therapy based on the detection of bacterial resistance before prescribing first-line therapy could help avoid the prescription of ineffective H. pylori eradication therapies and, overall, contribute to the control of antibiotic resistance of H. pylori.