Human adenoviruses (HAdVs) are common pathogens that are associated with a variety of diseases, including respiratory tract infections (RTIs). Without reliable, fast, and cost-effective detection methods for HAdVs, patients may be misdiagnosed and inappropriately treated. To address this problem, we have developed a multiplex loop-mediated isothermal amplification (LAMP) assay for the detection of the species Human adenovirus B (HAdV-B), Human adenovirus C (HAdV-C) and Human adenovirus E (HAdV-E) that cause RTIs. This multiplexing approach is based on the melting curve analysis of the amplicons with a specific melting temperature for each HAdV species. Without the need for typing of HAdVs, the LAMP results can be visually detected using colorimetric analysis. The assay reliably detects at least 375 copies of HAdV-B and -C and 750 copies of HAdV-E DNA per reaction in less than 35 min at 60 °C. The designed primers have no in silico cross-reactivity with other human respiratory pathogens. Validation on 331 nasal swab samples taken from patients with RTIs showed a 90–94% agreement rate with our in-house multiplex quantitative polymerase chain reaction (qPCR) method. Concordance between the quantitative and visual LAMP was 99%. The novel multiplexed LAMP could be an alternative to PCR for diagnostic purposes, saving personnel and equipment time, or could be used for point-of-care testing.

Острый респираторный дистресс-синдром (ОРДС) при COVID-19 отличается худшим прогнозом по сравнению с ОРДС при других заболеваниях. Летальность от ОРДС при COVID-19 составляет 26,0—61,5%, а обусловленная другими причинами, — 35,3—37,2%. ЦЕЛЬ ИССЛЕДОВАНИЯ Определение соотношения полиморфно-ядерных лейкоцитов (ПЯЛ), лимфоцитов и макрофагов в клеточном составе воспалительного инфильтрата в разные стадии и фазы диффузного альвеолярного повреждения (ДАП) при COVID-19 на материале аутопсий. МАТЕРИАЛ И МЕТОДЫ Исследована ткань легких 25 умерших от острой дыхательной недостаточности вследствие развития ОРДС при COVID-19 без вторичной бактериальной или микотической инфекции, а также при другой танатологически значимой патологии легких. Для изучения клеточного состава воспалительного инфильтрата и динамики его изменений использовали метод двойного иммуногистохимического исследования экспрессии антител к CD15, CD3 и CD68. РЕЗУЛЬТАТЫ Установлено, что воспалительный инфильтрат и интраальвеолярный экссудат в экссудативной фазе ДАП имеет следующий клеточный состав: 56,8% ПЯЛ (CD15-позитивные клетки; здесь и далее — среднее значение процентного отношения позитивных клеток к общему количеству клеток воспалительного инфильтрата), 6,9% лимфоцитов (CD3-позитивные клетки) и 19,5% макрофагов (CD68-позитивные клетки). При ранней стадии пролиферативной фазы выявлено 14,1% ПЯЛ, 38,7% лимфоцитов и 13,5% макрофагов; при поздней стадии пролиферативной фазы — 11,3% ПЯЛ, 14,5% лимфоцитов и 39,3% макрофагов. ЗАКЛЮЧЕНИЕ В экссудативной фазе ДАП выявлено статистически значимое преобладание ПЯЛ, что могло определять основной объем поражения легких и тяжесть течения ОРДС при COVID-19. Ранняя стадия пролиферативной фазы ДАП характеризуется статистически значимым изменением состава воспалительного инфильтрата по сравнению с экссудативной фазой в виде существенного уменьшения содержания ПЯЛ относительно общего количества клеток воспалительного инфильтрата; увеличением числа лимфоцитов, что, вероятно, связано с началом процессов организации и репарации. Поздняя стадия пролиферативной фазы ДАП по сравнению с ее ранней стадией отличается статистически значимым увеличением количества макрофагов в относительных показателях.

There are hundreds of coronaviruses, most of which circulate among animals, yet there are seven types that infect humans. Three of them can cause severe acute respiratory illness—SARS-CoV, SARS-CoV-2, and MERS-CoV. Other HCoV-229E, HCoV-OC43, HCoV-NL63, and HCoV-HKU1 usually cause only mild to moderate upper respiratory tract infections. These four coronaviruses are called seasonal, because they are continuously circulating among human population and are responsible for up to 30% of all respiratory tract infections. Genetically, these low-pathogenic types are related to SARS-CoV-2. That is why questions concerning the cross-reactivity and cross-neutralization between antibodies against different types of coronaviruses have been raised. We addressed these questions by using enzyme-linked immunosorbent assays and targeted next-generation sequencing (NGS). We established the upper respiratory infection etiology for three patients who had been vaccinated with Sputnik V and tested positive on anti-SARS-CoV-2 antibodies. The symptoms included sore throat, nasal congestion, and myalgia. Their blood serum was analyzed for anti-SARS-CoV-2 antibodies in dynamics: before vaccination, and after the first and second dose of the vaccine. After the second dose, all patients were positive for IgG antibodies against SARS-CoV-2. The targeted NGS panel sequencing data analysis showed that these patients were infected with common coronavirus HCoV-OC43. These results suggest that S protein-targeted vaccine-induced antibodies against SARS-CoV-2 are not protective against seasonal coronavirus HCoV-OC43.

The efficacy of immune checkpoint inhibitors (ICIs) in non-small-cell lung cancer (NSCLC) varies widely across patients. Growing evidence indicates that the gut microbiome, through its interaction with the tumor microenvironment, may influence the response to immunotherapy. To investigate this, we analyzed fecal and tumor samples from 63 patients with inoperable NSCLC undergoing ICI therapy. Based on microbiome profiling using 16S rRNA sequencing, patients were grouped according to treatment benefit, defined as progression-free survival (PFS) of six months or longer. Associations between α-diversity indices, microbial composition at the genus and phylum levels, and a composite Sum Index of Binary Abundance (SIBA) were examined in relation to clinical outcomes. Higher microbial α-diversity was linked to improved response to ICIs (p-value = 0.0078 for the Chao1 index). Multiple specific taxa, such as Ruminococcus gauvreauii (p-value = 2 × 10⁻⁴), Ruminiclostridium 9 (p-value = 8 × 10⁻⁴), and [Eubacterium] ventriosum (p-value = 9 × 10⁻⁴), were enriched in patients with favorable outcomes, whereas Oscillibacter and the Eubacterium hallii group were associated with disease progression (p-value = 2 × 10⁻³ and 9 × 10⁻³, respectively). The SIBA index, which reflects the absence of multiple beneficial bacterial taxa, proved to be a stronger predictor of treatment response than individual taxa alone. Median SIBA values were 18 vs. 24 in patients benefiting from IO therapy compared to non-responders (p-value = 9 × 10⁻⁷). These findings suggest that gut microbiome diversity and composition are closely tied to immunotherapy outcomes in NSCLC. Composite microbial metrics like SIBA may enhance predictive accuracy and inform personalized treatment approaches.