Для корреспонденцииЗулькарнаев Алексей Батыргараевичдоктор медицинских наук, профессор кафедры трансплантологии, нефрологии и искусственных органов факультета усовершенствования врачей ГБУЗ МО «Московский областной научно-исследовательский клинический институт им. М.Ф. Владимирского»,
The aim of the study was to evaluate the diagnostic value of the method of loop-mediated isothermal amplification of DNA with real-time fluorescent detection (real-time LAMP, or RT-LAMP) on the model of carnivore parvoviruses. Materials and methods. Samples of feces, blood and swabs from the rectum of different species of predatory animals with parvovirus enteritis (n = 39) and healthy animals (n = 31), as well as laboratory strains of mink enteritis virus, were analyzed by RT-LAMP using SYTO-9 and SYTO-82 dyes. Real-time PCR was used as a reference method. Results. In our study, the LAMP method with real-time fluorescence detection (RT-LAMP) in the carnivore parvovirus enteritis model provides high analytical sensitivity (1.5×103 copies of DNA/ml), diagnostic sensitivity and specificity (up to 100% under optimal conditions). Comparison of the two intercalating dyes showed that the SYTO-82 dye provides a higher signal-to-background ratio (22.6 ± 2.1) than the SYTO-9 dye (6.3 ± 1.5) (p <0.0000001 ). At the same time, SYTO-9 dye at the sensitivity limit (10 copies of DNA) provides an increase in fluorescence in the reaction mixture 13 minutes earlier than for SYTO-82 (23 and 36 minutes, respectively). Conclusion. RT-LAMP is a promising method for rapid and highly sensitive «point-of-care» diagnosis of infectious diseases, as well as in conditions of livestock farms or in field conditions.
Abstract. The article presents the results of a comparative assessment of the frequency of alleles and genotypes for the gene of beta-casein (β-CN) in cattle of the Kostroma and Yaroslavl breeds. The data on 137 bulls-producers of comparable and related breeds, presented on the official sites of “OOO Head Center for the Reproduction of Farm Animals” and the breeding farm “OOO Yaroslavskoe”, have been analyzed. The blood of cows from the Kostroma and Yaroslavl breeds in the breeding farms of the Kostroma region served as the object of DNA research. A total of 80 animals were genotyped. Evaluation of the distribution of the desired genotype β-CNА2/А2 in stud bulls showed that two breeds are in the lead – the Schwyz and the Kostroma breed, with the frequency of occurrence – 0.9090 and 0.6250, respectively. Moreover, in the Kostroma breed, there are no bulls carriers of the homozygous β-CNА1/А1 genotype. If we take into account the frequency of distribution of the β-CNА2 allele in bulls of the analyzed breeds, then it is higher – in the Schwyz breed – 0.9091, Kostroma – 0.8125 and Holstein – 0.6776. In bulls of the Yaroslavl breed there is 0.2083 frequency of the β-CNА2/А2 genotype and 0.4167 frequency of the β-CNА2 allele. A comparative assessment of the polymorphism data of the beta-casein locus in cows of the Yaroslavl and Kostroma breeds showed that in the Yaroslavl breed there were least of all animals with the β-CNА2/А2 genotype – 0.0333 and more – with the β-CNА1/А1 genotype – 0.6667. In the Kostroma breed, the genetic situation is completely different. So cows with genotypes β-CNА2/А2 – 0.5490, and with genotype β-CNА1/А1 – 0.0785. However, it is necessary to take into account the main indicators of cattle productivity in order to consider the beta-casein genotype as an additional selection trait. It is not possible to study the milk productivity of Yaroslavl breed cows with different genotypes, since only one cow with the CNА2/А2 genotype has been identified. However, cows of the Kostroma breed homozygous for the CNА2 allele, for the third and older lactations, had a milk yield of 1534 kg higher than cows homozygous for the CNА1 allele. Consequently, the beta-casein genotype can be considered an additional promising breeding trait.
Standard asthma therapy includes prescription of β2-agonists. Changes in the functional activity of β2-adrenergic receptor are associated with ADRB2 genepolymorphism and related to the low therapeutic response to β2-agonists. Identification of carriers of the clinically significant gene variants will help to avoidineffective treatment and prescribe an alternative therapy. This study aimed to assess clinical significance of the ADRB2 gene polymorphisms (Arg16Gly andGln27Glu) associated with the therapeutic response to β2-agonists in the group of asthma patients. We subjected a small group of adult nonsmoking patients(n = 21) with moderate asthma (III–IV stage of GINA) to clinical and genetic examination. The group included patients with the new theratype, those that poorlyrespond to β2-adrenergic drugs but significantly to M-cholinergic agonists. The first group included patients responding well to both salbutamol and ipratropiumbromide. The second group was comprised of the patients for whom salbutamol was not effective but who tested positive for response to ipratropium bromide. Theanalysis of distribution of polymorphic variants of Arg16Gly and Gln27Glu revealed no significant relationship between alleles and genotypes and the efficacy of β2-agonists(0.52 for the rs1042713 variant, p = 1.0; 1.0 for the rs1042714 variant, p = 0.74, respectively). The genotype of patients that did not respond to salbutamol waseither Arg16Gly or Gly16Gly. Further studies are needed that would involve a larger number of patients and an expanded list of the tested polymorphic variants.
Статья представляет собой реферативно-аналитический обзор исследований, посвященных изучению полиморфизмов генов 2-адренорецептора (ADRB2) и их ассоциаций со степенью тяжести бронхиальной астмы (БА) и ответа на стандартную фармакотерапию. Результаты анализа продемонстрировали высокую неоднородность полученных данных, что усугубляется различным целями, задачами и дизайном этих исследований. Учитывая то, что полиморфизм гена ADRB2 влияет на экспрессию данного рецептора, можно было бы предположить, что такие пациенты с БА будут демонстрировать низкий ответ на короткодействующие 2-агонисты (КДБА) и длительно действующие 2-агонисты (ДДБА), а в качестве бронхолитической терапии для них предпочтительны коротко- и длительно действующие холинергические средства. По изученным нами публикациям невозможно сделать однозначного вывода о связи полиморфизмов генов ADRB2 и ответа на КДБА и ДДБА. Данные, демонстрирующие распространенность гомозиготных аллелей и изучаемых ассоциаций среди представителей различных популяций, противоречивы. Достижение контроля над симптомами является основной целью терапии БА, в то же время, несмотря на наличие современных эффективных фармакологических средств, для ограниченного контингента среди пациентов с БА это не представляется возможным. Данные аспекты обусловливают необходимость изучения молекулярно-генетических механизмов патогенеза БА с целью улучшения диагностики и индивидуального подхода к выбору фармакотерапии для пациентов с плохим контролем БА.This article represents an abstract and analytical review of studies devoted to the polymorphisms of 2-adrenergic receptor gene (ADRB2) and their associations with the degree of severity of bronchial asthma (BA) and response to standard pharmacotherapy. The results of the analysis showed a high heterogeneity of the data obtained, which is aggravated by different goals, objectives and study design. If the polymorphism of ADRB2 gene affects the expression of this receptor, it is possible that these asthma patients will show a low response to SABA / LABA short- and long-acting anticholinergics will be preferred as bronchodilators. It is impossible to make a unique conclusion about the association between the polymorphisms of ADRB2 gene and the SABA / LABA responsiveness according to the results of analyzed publications. Data of the prevalence of homozygous alleles and studied associations among different populations are contradictory. Achievement of the asthma symptoms control is the main goal of asthma therapy. At the same time its impossible for a limited number of asthma patients in spite of the provision of current effective pharmacological resources. These aspects necessitate the research of the molecular-genetic mechanisms of pathogenesis in order to improve diagnostics and individual approach to the pharmacotherapy choice for patients with poor asthma control.
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