Extraction and Serological Properties of Mycobacterium Cell Surface and Excreted Proteins

Khaertynov, Kamil S.; Valeeva, Anna R.; Ivanov, Arkadiy V.; Mukminov, Malik N.; Urazov, Nail; Хаертынова, И. М.; Александрова, Н. М.; Moskvicheva, Albina V.; Ефимова, М. А.; Ахмадеев, Р. М.; Samigullina, Elvira S.; Nabatov, Alexey A.; Shuralev, Eduard A.

doi:10.1007/s12668-017-0492-1

Cited by 4 publications

(5 citation statements)

References 30 publications

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“…В данном исследовании мы использовали антиген, полученный из клеточных стенок M. bovis во фракциях после хроматографической очистки, в которых присутствовал отдельно антиген молекулярной массой 28 кДа (по результатам электрофореза в ПА-АГ) специфичный в вестерн-блоте с гипериммунной сывороткой крови кролика против цельного возбудителя туберкулёза. Модель антигена с молекулярной массой 28 кДа была взята за основу ввиду стандартизации получения данного антигена и его выраженных иммуногенных свойств, однако в полевых условиях необходимо применение мультиантигенного подхода [1,10,11]. Для нанесения на нитроцеллюлозную мембрану в нашем случае рабочая концентрация составила от 10 мг/мл, что существенно превышает расход антигена в ИФА даже несмотря на то, что объем наносимого антигена на одну мембрану (для сенсибилизации аналитической зоны нитроцеллюлозной мембраны шириной 5 мм, что соответствует одному готовому тесту для ИХА) составляет около 0,2-0,3 мкл.…”

Section: результаты / Resultsunclassified

“…Для выделения антигенов микобактерий туберкулёза (M. bovis, штамм bovinus 8 -700201), культивировали на питательной среде Левенштейна-Йенсена в течение 1-2 месяцев -до появления видимого роста бактерий, не допуская их перерастания, после чего выделяли антигены [4].…”

Section: материалы и методы / Materials and Methodsunclassified

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An immunochromatographic test for the detection of anti-tuberculosis antibodies

Khammadov,

Khaertynov,

Akhmadeev

et al. 2023

jour

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Tuberculosis is a dangerous socially significant disease of various animal species. According to Rosselkhoznadzor data for 2022, the situation with bovine tuberculosis is endemic, stable, long-term trends are decreasing, epidemic thresholds for ill health and incidence have not been overcome. It should be noted that in 2022, bovine tuberculosis was not officially detected, but this infection was registered in several pigs and wild boars. Take a point that the reduction in the number of cases of tuberculosis in cattle is decreased (to zero), the overall strategy for combating this disease is successful, but this does not mean that the need to develop new tests for the accelerated diagnosis of this infection has completely disappeared. The safety of service personnel directly depends on minimizing the risk of infection with pathogens common to humans and animals, one of which is tuberculosis. The fastest way to indicate infectious agents is by immunochromatographic analysis. This publication is devoted to the details that it is desirable to focus on when developing a diagnostic test based on immunochromatographic analysis, using the example of bovine tuberculosis. As an antigen in the presented study, native antigens of Mycobacterium bovis grown on a nutrient medium Levenshtein-Jensen with proven antigenic activity were used. Conjugation of antispecies antibodies was carried out with a colloidal solution of gold, (diameter of particle is 25±0,9 nm). In the course of the work, the optimal conditions for the manufacture of a diagnostic test for the detection of anti-tuberculosis antibodies by immunochromatographic analysis were shown.

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Section: результаты / Resultsunclassified

Section: материалы и методы / Materials and Methodsunclassified

An immunochromatographic test for the detection of anti-tuberculosis antibodies

Khammadov,

Khaertynov,

Akhmadeev

et al. 2023

jour

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“…Antigens were isolated from M. bovis Bovinus-8 strain 700201 (obtained from the Scientific Center for Drug Expertise of the Ministry of Health of the Russian Federation) according to the previously described methodology [20]. Cells were cultured on solid Lowenstein-Jensen medium for 28-30 days at 37°C, then destroyed using FastPrep-24 homogenizer with Lysing MatrixB (MP Biomedicals, USA).…”

Section: Extraction Purification and Characterization Of Mycobacteria...mentioning

confidence: 99%

Effect of Isolated <em>Mycobacterium bovis</em> Proteins on the Respiratory Response of Mouse Granulocytes

Gabdoulkhakova,

Korovina,

Filina

et al. 2023

Preprint

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Proteins are considered to be the main carriers of antigenic properties of Mycobacterium tuberculosis and Mycobacterium bovis, the causative agents of tuberculosis, the most serious infectious disease. Neutrophils, following resident macrophages, are among the first to attack the mycobacteria in defense of the organism. Neutrophils themselves are also the target of mycobacterial products; and their effect on neutrophil functions has been less studied. We have obtained about 70 protein fractions from M. bovis lysate; among them four protein fractions (10, 22, 24, and 40 kDa) had high serological activity against the serum of a rabbit immunized with M. bovis cells. We studied the effect of the serologically active proteins isolated from M. bovis on the generation of reactive oxygen species by bone marrow granulocytes of the Balb/c mouse, evaluated by luminol-dependent chemiluminescence. For comparison, two synthetic peptides (both 3 kDa) carrying immunogenic epitopes of MPB 70 protein (M. bovis) were taken. Proteins of 10, 22, and 24 kDa at concentrations of 83, 80, and 59 ng/mL, respectively, were found to activate ROS generation by mouse granulocytes. The 10- and 40-kDa antigens inhibited the receptor-mediated respiratory responses to opsonized zymosan (OZ) and N-formyl-MLF peptide (fMLF) but significantly enhanced the response to the phorbol ester PMA, which directly activates protein kinase C. Antigens with molecular weights of 22 and 24 kDa enhanced the fMLF-induced response and had no effect on the response activated by PMA. The response to OZ was suppressed in the presence of the 22 kDa protein. Synthetic peptides over a wide range of concentrations showed no activity in any case. All M. bovis proteins examined did not affect both proliferation and cell viability of the A549 and THP-1 cells; therefore, they have no cytotoxic or proliferative effects. Thus, serologically active proteins with molecular weights of 10, 22, 24, and 40 kDa isolated from M. bovis modulated the respiratory responses of mouse granulocytes induced by the formyl peptide, while proteins of 10, 22, and 40 kDa inhibited the responses induced by OZ.

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“…It is abundant on the surface of MTB, so it can be used as a simulant of MTB. 29,30 We performed surface functionalization on the prepared sensor. First, the surface of the sensor was treated with oxygen plasma, and then the above packaging method was used to encapsulate the microfluidic chip biosensor.…”

Section: Preparation Of Biosensors and Integration Of Microfluidic Chipsmentioning

confidence: 99%

Rapid detection of airborne protein from Mycobacterium tuberculosis using a biosensor detection system

Jiang

et al. 2022

Analyst

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Extraction and Serological Properties of Mycobacterium Cell Surface and Excreted Proteins

Cited by 4 publications

References 30 publications

An immunochromatographic test for the detection of anti-tuberculosis antibodies

An immunochromatographic test for the detection of anti-tuberculosis antibodies

Effect of Isolated <em>Mycobacterium bovis</em> Proteins on the Respiratory Response of Mouse Granulocytes

Rapid detection of airborne protein from Mycobacterium tuberculosis using a biosensor detection system

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