جستجوی مقالات مرتبط با کلیدواژه "mycobacterium bovis" در نشریات گروه "پزشکی"
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Background and Objectives
Lipoarabinomannan is one of the components of the significant structural cell surfaces of my- cobacteria and serves as an immunostimulatory factor. TNF-α and IL-12 are two examples of the anti-bacterial inflammatory cytokines that are activated and induced during infection.
Materials and MethodsIn this study, mannan was extracted and processed, and then Bulb/c female mice were used in three groups, one group was given BCG vaccine, the other group was given BCG vaccine with mannan adjuvant, and a non-injected group was used as a control group. Inflammatory factors interleukin-12, TNF-α, IgG and IgM were measured in mouse serum.
ResultsThe levels of the inflammatory factors interleukin-12 and TNF-α in the serum isolated from mice receiving the BCG vaccine with mannan adjuvant showed a significant difference compared to the group that received only the BCG vaccine and the control group [IL-12] and , with P≤0.05.The examination of the level of IgG immune factors in these three groups revealed a significant difference. The group that received the BCG vaccine with mannan adjuvant showed a marked contrast compared to the group that received only the BCG vaccine and the control group, with P≤0.05. The level of IgM was higher in the group that received the BCG vaccine alone compared to the adjuvant vaccine group and the control group, with P≤0.05.
ConclusionOur results indicated that mice receiving the BCG vaccine with mannan adjuvant had significantly higher serum levels of IL-12, TNF-α, and IgG than the group receiving BCG alone.
Keywords: Mycobacterium Bovis, Adjuvant, Mannan, Immunity -
Biolmpacts, Volume:12 Issue: 5, Sep 2022, PP 395 -404Introduction
The limited efficacy of BCG (bacillus Calmette–Guérin) urgently requires new effective vaccination approaches for the control of tuberculosis. Poly lactic-co-glycolic acid (PLGA) is a prevalent drug delivery system. However, the effect of PLGA-based nanoparticles (NPs) against tuberculosis for the induction of mucosal immune response is no fully elucidated. In this study, we hypothesized that intranasal immunization with culture filtrate protein-10 (CFP10)-loaded PLGA NPs (CFP10-NPs) could boost the protective immunity of BCG against Mycobacterium bovis in mice.
MethodsThe recombinant protein CFP10 was encapsulated with PLGA NPs to prepare CFP10-NPs by the classical water–oil-water solvent-evaporation method. Then, the immunoregulatory effects of CFP10-NPs on macrophages in vitro and on BCG-immunized mice in vivo were investigated.
ResultsWe used spherical CFP10-NPs with a negatively charged surface (zeta-potential −28.5 ± 1.7 mV) having a particle size of 281.7 ± 28.5 nm in diameter. Notably, CFP10-NPs significantly enhanced the secretion of tumor necrosis factor α (TNF-α) and interleukin (IL)-1β in J774A.1 macrophages. Moreover, mucosal immunization with CFP10-NPs significantly increased TNF-α and IL-1β production in serum, and immunoglobulin A (IgA) secretion in bronchoalveolar lavage fluid (BALF), and promoted the secretion of CFP10-specific interferon-γ (IFN-γ) in splenocytes of mice. Furthermore, CFP10-NPs immunization significantly reduced the inflammatory area and bacterial load in lung tissues at 3-week post-M. bovis challenge.
ConclusionCFP10-NPs markedly improve the immunogenicity and protective efficacy of BCG. Our findings explore the potential of the airway mucosal vaccine based on PLGA NPs as a vehicle for targeted lung delivery.
Keywords: PLGA, Nanoparticles, CFP10, Mycobacterium bovis, Mucosal immunization -
Bacillus Calmette Guerin (BCG) was designed for protecting children against tuberculosis. Also, it can protect against other infectious diseases through the induction of trained immunity. Due to its heterologous protective effects, the BCG vaccine has been proposed as a treatment option for coronavirus disease-2019 (COVID-19). Epidemiological studies have found that countries without BCG vaccination policy have experienced higher mortality rates related to COVID-19 infection than those with BCG vaccination policy. However, there are some confounding factors such as age, population intensity, immigration, the pandemic phase, and data accuracy that may affect these results. Therefore, this hypothesis should be evaluated by clinical trial studies. Large-scale clinical trials are in progress to investigate if the BCG vaccine could be used as a useful tool for protection against COVID-19 infection.
Keywords: COVID-19, Mycobacterium bovis -
Bacillus Calmette–Guérin (BCG) has been used as an intravesical product for the treatment of intermediate and high risk, non-muscle-invasive bladder cancer (NMIBC). Freeze drying technique is highly recommended for product development, however, the microorganism sensitivity to freezing and drying processes is a major chalenge which may lead to poor survival. To overcome this problem, the use of cryoprotectants in intravesical BCG formulation is required. This study was, therefre, planned to design a new formulation, using an attenuated strain of Mycobacterium bovis, which could be produced by freeze drying technique with the aim of prolonging its storage stability and increasing its efficacy as well as the ease of administration. For this purpose, sodium L-glutamate monohydrate (a commonly used stabilizer in domestic BCG suspension formulations) was replaced by lactose monohydrate. New intravesical BCG formulations, both in lyophilized and liquid forms, were eventually evaluated by moisture content assay, viable count assay, bacterial and fungal contamination, safety test and determination of bacterial concentration and O2 consumption. The results were compared with the data obtained for the conventional lyophilized and liquid products. Maximum survival rate was achieved in the presence of 10 % w/v lactose monohydrate for both liquid and lyophilized formulations when stored at less than -10 and 2-8C, respectively. In summary, the freeze-dried formulations developed with lactose monohydrate met the requirements of intravesical BCG in high viability and stability during storage.
Keywords: Bacillus Calmette–Guérin, Cryoprotectant, Freeze drying, Intravesical BCG, Lactose monohydrate, Mycobacterium bovis -
Introduction
The diagnosis of Tuberculous Meningitis (TBM) has remained a challenge due to its insidious onset and the failure of conventional diagnostic tests. The present study aimed to identify the mycobacterial pathogen in the CSF of patients with TBM and a poor prognosis.
MethodsWe retrospectively recruited 224 TBM and 34 non-TBM patients admitted to the Central India Institute of Medical Sciences, Nagpur, India, in 2014. The CSF samples of these patients were subjected to a duplex PCR assay for the species-specific identification of the causative pathogen.
ResultsM. bovis and infection with M.tuberculosis were detected in 7% (18) and 32.9% (85) of the patients, respectively. Moreover, 14% (36) of the study samples were culture positive; however, the mycobacterial pathogens could not be differentiated to the species level.
ConclusionThe present study findings emphasized the potentially vital importance of M. bovis identification for appropriate patient management. The obtained data also demonstrated the persistent significance of M. bovis, as a zoonotic pathogen.
Keywords: Duplex PCR, Mycobacterium Bovis, Tuberculous meningitis, Zoonoses -
Background and Objectives
Bovine tuberculosis diagnosis is usually performed by various tests with specific limitations. Mycobacterium bovis culture filtrate contains antigenic proteins that could be used to improve the sensitivity of bovine tuberculosis diagnosis. The objective of this study was to identify and purify antigenic proteins from culture filtrates of M. bovis strain AN5 for use in immunological assays.
Materials and MethodsSecreted proteins were purified from the heat-treated culture filtrate of M. bovis strain AN5. Proteins were precipitated with ammonium sulfate, fractionated by Sephadex G50 chromatography. The protein concentrations and the approximate molecular weight were determined by lowry method and 12% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), respectively. Immunological methods, including dot-blotting and western blotting, assessed the quality of the isolated proteins.
ResultsThe quantity of antigenic proteins in the culture medium was measured at far more than 15% of the amount of proteins secreted into medium. Three main chromatographic fractions obtained and showed concentrations of proteins ranging from 14 to 60 μg/μl with molecular weights in the 10 to 180 kDa range. The purified antigens showed positive reactions to the infected cattle serum throughout dot-blotting. Western blotting revealed a total of 15 to 70 kDa molecular weight proteins.
ConclusionImmunoblotting analysis made it possible to detect and recognize novel antigens that are useful for bovine tuberculosis diagnosis improvement. This is significant since non-specific reactions were not observed when we utilized serum of cattle experimentally infected with M. bovis as a polyclonal antibody.
Keywords: Mycobacterium bovis, Purified antigenic proteins, Dot-blot, Western blot -
زمینه
عامل اصلی سل در گاو، مایکوباکتریوم بوویس است. متداول ترین روش در حال استفاده جهت شناسایی گاوهای آلوده، در مقیاس جهانی و همچنین در ایران، تست توبرکولین است.
هدفمطالعه حاضر با هدف بهبود دانش ما از ساختار جمعیت مایکوباکتریوم بوویس های جدا شده در گاوداری های شیراز صورت پذیرفت.
مواد و روش هادر این مطالعه مقطعی توصیفی که بین دی 1394 تا بهمن 1395 انجام شد، 50 نمونه پاتولوژیک جمع آوری شده از گاوهای توبرکولین مثبت، از دو کشتارگاه در شیراز، بر روی محیط لونشتاین جانسون گلیسیرینه و پیرواته کشت داده شدند. استخراج ژنوم از کشت های مثبت انجام شد و از آن ها در آزمون های PCR-16SrRNA ،PCR-IS6110، و PCR-RD Typing استفاده شد. تمام جدایه های مایکوباکتریوم بوویس با استفاده از هضم آنزیمی با آنزیم pvu II و روش PGRS-RFLP مورد ژنوتایپینگ قرار گرفتند.
یافته هادر کشت باکتریایی، درمجموع 13 نمونه (26 درصد) حاوی مایکوباکتریوم زنده بودند که با استفاده از آزمون های PCR، هویت همه جدایه ها به عنوان مایکوباکتریوم بوویس تایید شد. ژنوتایپینگ با استفاده از روش PGRS-RFLP، دو الگو را نشان داد که 10 جدایه ژنوتیپ مشابه و سه جدایه ژنوتیپ متفاوتی با سویه مایکوباکتریوم بوویس GCB (1173 P2) داشتند.
نتیجه گیریدر حالی که تداوم مایکوباکتریوم بوویس شبه BCG (به عنوان یک ویژگی معمول در جمعیت مایکوباکتریوم بوویس ایرانی) در گاوداری های شیراز تعجب آور نیست، ممکن است که شناسایی یک سویه بسیار مشابه در کار ما بتواند نشان دهنده تکامل موضعی سویه های جدید مایکوباکتریوم بوویس در منطقه یا نفوذ چنین سویه هایی از طریق فعالیت های دامپروری باشد.
کلید واژگان: مایکوباکتریوم بوویس, تست توبرکولین, PGRS-RFLP, PVU II, سویهBackgroundMycobacterium bovis is the main cause of tuberculosis in cattle. The most commonly used method to identify bovis-infected cattle is tuberculin test.
ObjectiveThe present study aimed to investigate the population structure of Mycobacterium bovis in infected cattle farms of Shiraz City in Iran.
MethodsIn this descriptive cross-sectional study, 50 pathological samples from tuberculin-positive cattle that were collected from two abattoirs were cultured on glycerinated and pyruvated Lowenstein-Jensen media. Genomic material from culture-positive slopes was extracted and used in polymerase chain reaction (PCR)-16S rRNA, PCR-IS6110, and PCR--regions of difference (RD) typing. All the M. bovis isolates were then digested by PvuII restriction enzyme and genotyped by polymorphic guanine/cytosine-rich repetitive sequences (PGRS)-restriction fragment length polymorphism (RFLP) technique.
FindingsIn bacterial culture, 13 (26%) of samples had living mycobacteria where PCR test results revealed their identity as Mycobacterium bovis. Genotype profiling by RFLP-PGRS method displayed two patterns with 10 isolates shared a single profile identical to that of M. bovis bacillus calmette-guerin (BCG) strain (1173 P2) and three isolated with a different genotype.
ConclusionHigher prevalence of BCG-like M. bovis (as a typical characteristic of Iranian M. bovis population) in cattle farms of Shiraz City was expected. This may indicate the local evolution of new M. bovis strains in the region or the infiltration of such strains through cattle farming activities.
Keywords: Mycobacterium bovis, Tuberculination, PGRS-RFLP, PVUII, Strain -
Thiosemicarbazides are potent intermediates for the synthesis of pharmaceutical and bioactive materials and thus, they are used extensively in the field of medicinal chemistry. The imine bond (-N=CH-) in these compounds are useful in organic synthesis, in particular for the preparation of heterocycles and non-natural β-aminoacids.In this paper the synthesis of some new thiosemicarbazide derivatives by condensation reaction of various aldehydes or ketones with 4-phenylthiosemicarbazide or thiosemicarbazide is reported. This synthesis method has the advantages of high yields and good bioactivity. The structures of these compounds were confirmed by IR, Mass, 1H NMR, 13C NMR, and single-crystal X-ray diffraction studies. All of these compounds were tested for their in vitro anti-mycobacterial activity. The influence of the functional group and position of substituent on anti-bacterial activity of compounds is investigated too. The preliminary results indicated that all the tested compounds showed good activity against the test organism. The compounds 11 and 30 showed the highest anti-tubercular activity (0.39 μg/mL).Keywords: Thiosemicarbazide derivatives, Biological evaluation, Mycobacterium bovis, anti-tubercular activity, Medicinal chemistry
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A series of new fluorene bisamide derivatives were synthesized through multi-component Ugi reaction and tested for their in vitro anti-mycobacterial activity. The structures of the products 5a-w were deduced from their IR, 1H NMR, and 13C NMR spectra. Elemental analyses (CHN) for novel compounds (5a, 5d, 5f, 5h, 5k, 5l, 5p, 5s, 5t, 5v, 5w) was done to. These compounds were evaluated as anti-bacterial agents against Mycobacterium bovis and M. tuberculosis, while their activity expressed as the minimum inhibitory concentration (MIC) in μg/mL. Among the twenty-three synthesized compounds, 5a was found to be the most active compound in vitro with MIC of 1.95μg/mL against Mycobactrium bovis and compound 5k showed greatest potency against sensitive and resistant strains of M. tuberculosis (H37Rv, IHMT149/09, HPV115/08 and HPV65/08).Keywords: multi-component reaction, Ugi reaction, Mycobacterium bovis, fluorene bisamide derivatives, brine shrimp toxicity study
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IntroductionThe Bacille Calmette-Guérin (BCG), a live attenuated Mycobacterium bovis vaccine, is administered to all the newborns in Tunisia in order to prevent Tuberculosis (TB). Complications of this vaccine are uncommon. However, it poses a risk for children with unknown immunodeficiency.Case PresentationWe report on disseminated BCG disease in two infants, respectively, with severe combined immunodeficiency and human immunodeficiency virus (HIV). Evolution was fatal for both, despite adequate anti-tuberculosis treatment.ConclusionsMolecular methods are available to respond to the urgent need for rapid and specific diagnosis of local/regional or systemic BCG disease, using available commercial kits GenoType® MTBC and GenoType® MTBDRplus. These tests allow prevention of inoculation of live vaccines such as BCG among the next siblings until appropriate screening tests exclude primary or secondary immunodeficiency syndromes.Keywords: Bacille Calmette, Guérin Vaccine, Mycobacterium bovis, Molecular Diagnostic Techniques, Immunologic Deficiency Syndromes
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زمینه و هدفبیماری سل، از شایع ترین بیماری های عفونی در جهان است که سازمان جهانی بهداشت، آن را به عنوان یک اورژانس جهانی اعلام کرده است. با وجود شناخت عامل این بیماری و وجود داروهای موثر برای درمان آن، هنوز این بیماری یک معضل جهانی است. هدف از این مطالعه جدا سازی و تعیین هویت سویه های کمپلکس مایکوباکتریوم توبرکلوزیس با روش PCR-RD Typing در مسلولین استان زنجان و شناسایی موارد سل زئونوز می باشد.روش بررسیدر این مطالعه تعداد 27 نمونه کشت مثبت سال 1392 از مرکز بیماری های ریوی و سل استان زنجان دریافت شد و روی محیط های کشت لوین اشتاین جانسون گلیسیرین دار و پیروات دار مجددا تکثیر گردید. DNA جدایه ها به روش Van Soolingen استخراج و برای تایید جنس مایکوباکتریوم آزمون PCR-16SrRNAو تعلق به گروه کمپلکس مایکوباکتریوم توبرکلوزیس PCR-IS6110 و تعیین گونه توسط PCR- RD Typing (RD1، RD4، RD9، RD12)انجام شد.یافته هااز 27 جدایه، تمامی جزء جنس مایکوباکتریوم و متعلق به کمپلکس مایکوباکتریوم توبرکلوزیس بودند که 25 جدایه مایکوباکتریوم توبرکلوزیس و 2 جدایه مایکوباکتریوم بویس بودند.نتیجه گیرینتیجه ی به دست آمده از این مطالعه نشان می دهد که گونه های مایکوباکتریوم بویس در مسلولین استان زنجان وجود دارد و دلیلی بر زئونوز بودن بیماری سل و انتقال آن از حیوان به انسان می باشد و این زنگ خطری برای گسترش سویه های حیوانی در جامعه می باشد. برای همین در مواقع کار بر روی نمونه های انسانی مشکوک به سل، باید مسئله ی جداسازی سویه های زئونوز قابل انتقال از حیوان به انسان را مد نظر قرار داد.کلید واژگان: کمپلکس مایکوباکتریوم توبرکلوزیس, مایکوباکتریوم بویس, مایکوباکتریوم توبرکلوزیس, PCR, RD TypingBackground And ObjectiveTuberculosis (TB) is amongst the common infectious diseases in the world so that the World Health Organization (WHO) has declared it as a global emergency as well as an extremely contagious disease. Although the cause of the disease and its therapy are well known, TB is considered as a global dilemma. The present study was set to isolate and identify Mycobacterium Tuberculosis Complex by PCR-RD typing from TB patients in Zanjan province and to verify cases of zoonotic tuberculosis.Materials And MethodsIn this study, 27 positive samples were collected from center of pulmonary diseases and tuberculosis in Zanjan Province during 2013 and were grown on LowensteinJensen medium (LJP and LJG). DNA was extracted from the bacterial isolates by Van Solengen method and at genus level identification was done by PCR-16SrRNA. To confirm the mycobacterium tuberculosis complex, PCR-IS6110 was used. Identification of species was performed by PCR-RD typing (RD1, RD4, RD9 and RD12).ResultsA total of 27 isolates were obtained which were identified as genus Mycobacterium belonging to Mycobacterium tuberculosis complex. Twenty five of the isolates were Mycobacterium tuberculosis while 2 isolates were identified as M.bovis.ConclusionThe results of the study indicates that M.bovis species is present among the human TB cases in Zanjan province, which indicates the zoonotic potential of the species and the possible transmission of the species from animals to humans. These results might be alarming owing to the spread of the bovine strains among the human subjects. Thus, it is essential that during research on suspected human TB cases, the zoonotic species capable of transmission from animal to human be considered.Keywords: Mycobacterium tuberculosis complex, Mycobacterium bovis, Mycobacterium tuberculosis, PCR, RD typing
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مجله دانشکده پزشکی دانشگاه علوم پزشکی تهران، سال هفتاد و چهارم شماره 10 (پیاپی 190، دی 1395)، صص 675 -681با وجود پیشرفت های بسیار در حوزه دارو و واکسن در طی یک قرن گذشته بیماری توبرکلوزیس هنوز مسئول بیش از دو میلیون مرگ در هر سال می باشد. پیدایش سویه های مقاوم به دارو و عفونت همزمان توبرکلوزیس و ایدز آن را به یک مشکل جدی بدل کرده است و درمان و کنترل آن بار بزرگی بر دوش جامعه ایجاد نموده است. عفونت توبرکلوزیس در کودکان با عوارض شدیدی همراه است به ویژه در آن هایی که کمتر از پنج سال سن دارند، بنابراین تجویز واکسن به این دسته از افراد لازم است. باسیل کالمت-گورین (Bacillus Calmette-Guérin) سویه ضعیف شده ی مایکوباکتریوم بوویس بوده که برای اولین بار جهت ایجاد مصونیت بر ضد عفونت توبرکلوزیس مورد استفاده قرار گرفت. همچنین این واکسن توانایی ایجاد مصونیت نسبی بر ضد عفونت لپروزی، اولسربرولی بوده و همچنین جهت درمان سرطان مثانه نیز استفاده می شود. واکسن ب ث ژ تنها باید به افرادی با سیستم ایمنی سالم تزریق شده و از تجویز آن به افراد با سیستم ایمنی تضعیف شده باید اجتناب کرد و همچنین این واکسن در افرادی که پیش تر به عفونت توبرکلوزیس مبتلا شده اند و یا با مایکوباکتریوم های محیطی آلوده شده اند، اثربخش نمی باشد. مطالعات متاآنالیز گذشته نشان می دهد که میزان اثربخشی این واکسن بر ضد عفونت تنفسی متغیر بوده و بین 80%-0 می باشد اما بر ضد اشکال بسیار شدید منتشره مانند مننژیت و سل ارزنی بسیار کارآمد می باشد. با وجود بسیاری از تلاش ها و طراحی نسل جدیدی از واکسن ها بر ضدتوبرکلوزیس، واکسن ب ث ژ هنوز تنها واکسن موثر بر ضد عفونت توبرکلوزیس می باشد چرا که جایگزینی این واکسن با یک واکسن موثرتر موفقیت آمیز نبوده است.کلید واژگان: ب ث ژ, مایکوباکتریوم بوویس, لپروزی, سرطان مثانهBackgroundDespite advances in the vaccinology and chemotherapy in the past century, tuberculosis is still responsible for two million deaths every year. Emergence of multi-drug resistant strain and coinfection of TB-HIV make it a serious concern. Treatment and control of tuberculosis is a great health burden in every community. Active tuberculosis in children has very severe consequences especially those who are under 5-years-old, therefore vaccine indication should be taken. Bacille Calmette-Guérin (BCG) is a live attenuated strain of Mycobacterium bovis that has been used for providing immunity or protection against tuberculosis (TB). In addition, BCG provides relative protection against leprosy and Buruli ulcer, it also can be used for treatment of bladder cancer. BCG is the most widely administered vaccine around the world. It has been given to over three billion individuals over the past decades. At first it was developed in 1908 at the Pasteur Institute in Lille by Albert Calmette and Camille Guérin. In fact BCG is a strain of Mycobacterium bovis that bear deletion in its genome following too long subculture in special media. Deletion in region of deletion 1 (RD1), a specific region of Mycobacterium bovis genome, has decreased pathogenicity of BCG strain. Following culture of BCG on different media since 1921 make genetic variation in the BCG strains that have specific characteristics. BCG should begin given to only immune-competent individuals and should not be administered to immunocompromised people. This vaccine is not effective in people formerly infected or sensitized with environmental mycobacteria. Previous meta-analysis studies indicate that BCG has variable range of protection from 0 to 80 percent against pulmonary TB, but is very effective against severe disseminated forms such as meningitis and miliary form of TB. Despite many research and develop new generation vaccine against TB, BCG vaccine still remains as the only effective vaccine because many efforts to replace it with better ones were unsuccessful.Keywords: BCG vaccine, buruli ulcer, leprosy, mycobacterium bovis, neoplasms, urinary bladder
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At present, most of Iran is free of bovine tuberculosis (TB). The strategy of control and eradication in Iran involves a tuberculation test and slaughter of reactors, a procedure transformed the present-day prevalence of TB into a sporadic occurrence. This paper describes the first report of bovine tuberculosis in a European fallow deer (Dama dama dama) in Iran. The deer was emaciated and found dead in the Hoveize Provincial Zoo Park. Post-mortem examinations revealed multifocal granulomatous and suppurative abscesses in the lungs and mesenteric lymph nodes. These post-mortem indicators led the authors to suspect TB, and the PCR test and bacteriology tests confirmed it as an infection by the Mycobacterium bovis. This survey discusses the important implications of such findings for wildlife, especially livestock, as well as for human TB disease control, because deer are often conserved in public zoos and humans often come into contact with them.Keywords: Mycobacterium bovis, Tuberculosis, European fallow deer, Iran
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IntroductionThere are increasing reports of serious adverse events of bacillus Calmette-Guerin (BCG) vaccination in infants with unrecognized primary immunodeficiency disorders (PIDs) in our country. Among these adverse events skin manifestations occur less frequently and are less noticed..Case PresentationWe report on an 11-months-old boy with prolonged fever and diffuse hyper pigmented subcutaneous nodules. Due to lymphopenia, oral thrush and severe adverse reaction to BCG vaccination, the possibility of primary immunodeficiency was considered for him and immunological investigations were done..ConclusionsSubcutaneous nodules in the absence of a local reaction at the site of BCG vaccination may be the sole manifestation of disseminated BCG disease..Keywords: Mycobacterium Bovis, Skin, Immunologic Deficiency Syndromes
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فصلنامه علوم پزشکی دانشگاه آزاد اسلامی، سال بیست و پنجم شماره 2 (پیاپی 80، تابستان 1394)، صص 119 -124سابقه و هدفMycobacterium bovis به عنوان عامل سل گاوی به صورت موردی انسان را درگیر می کند و همراه با Mycobacterium tuberculosis به عنوان معضلات بهداشتی با گستره جهانی محسوب می شوند. هدف از انجام این مطالعه جداسازی و مقایسه پروفایل پروتئینی این دو سویه به منظور دستیابی به بیومارکر موثر در تشخیص و ایمونیزاسیون در مطالعات آینده بود.روش بررسیدر ابتدا نمونه های بالینی با روش ان استیل- ال سیستئین-هیدروکسید سدیم و در محیط کشت لونشتاین جانسون کشت داده شدند و جهت افتراق سویه ها از سایر مایکوباکتریوم ها، از تست های بیوشیمیایی و حساسیت آنتی بیوتیکی استفاده شد. کلونی ها در محیط میدل بروک 7H9 کشت داده شدند، سپس به منظور استخراج پروتئین های ترشحی و غشایی از روش های سونیکاسیون، رسوب دهی با سولفات آمونیوم و الکل استفاده گردید و به وسیله روش برادفورد تعیین غلظت شد و در نهایت مقایسه با روش الکتروفورز تک بعدی انجام پذیرفت.یافته هااز تفاوت های عمده باندهای حاصل از تفکیک پروتئین های غشایی بین دو سویه M. tuberculosis و M. bovis می توان به باندهای 45 و 60 کیلودالتونی و همچنین نواحی بین باندهای 14 تا 45 کیلودالتون در تفکیک حاصل از پروتئین های ترشحی بین دو سویه اشاره کرد.نتیجه گیریبه نظر می رسد اختلاف بین باندهای پروتئینی سویه های حساس و بوویس می تواند به عنوان پروتئین مارکر و یا حتی بیومارکر موثر در تشخیص سویه های توبرکلوزیس حساس و بوویس از یکدیگر قابل استفاده باشد و حتی از شباهت ها نیز می توان در ایمونیزاسیون استفاده نمود.
کلید واژگان: Mycobacterium tuberculosis, Mycobacterium bovis, SDS, pageMedical Science Journal of Islamic Azad Univesity Tehran Medical Branch, Volume:25 Issue: 2, 2015, PP 119 -124BackgroundMycobacterium bovine, as the etiology of bovine TB, involves human in some cases and with Mycobacterium tuberculosis are regarded as global health problems. The aim of this study was to extract and compare protein profile of these strains in order to achieve effective biomarkers for diagnosis and the vaccine components.Materials And MethodsFirst, the clinical samples were cultured on Lowenstein-Jensen (LJ) medium by the N-acetyl-L-cysteine-sodium hydroxide method and the biochemical tests and antibiotic susceptibility were used. Colonies were grown in the Middlebrook 7H9 medium and, upon harvesting the new colonies, secretory and membrane proteins were extracted by sonication and ammonium sulfate and alcohol precipitation. Concentration was determined by Bradford method and finally the comparison was made through one dimensional electrophoresis.ResultsThe major discrepancy between two strains of Mycobacterium tuberculosis and Mycobacterium bovis was 45 and 60 KDa bands as well as the zone between 14 and 45 KDa bands of secretory proteins, in the banding separation membrane proteins.ConclusionDiscrepancy in the protein bands of sensitive strains and Mycobacterium bovis seems to be used as an effective protein marker or even a biomarker in distinguishing the sensitive and bovis strains; also similarities can be used for immunization purposes. Keywords:Keywords: Mycobacterium tuberculosis, Mycobacterium bovis, SDS, page. -
سابقه و هدف
عفونت گوش میانی یکی از بیماری های شایع در میان کودکان است. عوامل مختلفی از جمله باکتری ها و ویروس ها عامل ابتلا به عفونت گوش میانی هستند. شایع ترین باکتری های پاتوژناسترپتوکوکوس پنومونیه، هموفیلوس آنفلوانزا، موراکسلاکاتارالیس و استرپتوکوکوس پی وژن هستند. به تازگی الویوکوکوس اوتیتیدیس به عنوان یکی از عوامل ایجاد کننده این عفونت شناخته شده است که مطالعه های محدودی به دلیل سخت رشد و کند رشد بودن و شرایط سخت نمونه گیری روی این باکتری صورت گرفته است. هدف از انجام این مطالعه تعیین فراوانی و جداسازی الویوکوکوس اوتیتیدیس از ترشحات گوش میانی کودکان مبتلا به اوتیت میانی به روش کشت و PCR است.
موادوروش هاتحقیق به روش توصیفی انجام و تعداد 60 نمونه از ترشحات عفونت گوش میانی از سوی پزشک متخصص جمع آوری شد. از میان نمونه های اتخاذ شده بخشی از حجم مایع گوش میانی برای کشت روی محیط کشت میکروبی تلقیح و بخشی دیگر برای انجام کارهای مولکولی استفاده شد. عفونت گوش با دو روش بررسی و با آزمون کای دو مورد قضاوت آماری قرار گرفت.
یافته هااز تعداد 60 نمونه عفونت گوش میانی تعداد 3 (5درصد)نمونه به روش کشت شناسایی شد و با روش PCR 28 (46درصد) نمونه مثبت ارزیابی شدند(P<0/0001).
نتیجه گیریبه نظر می رسد الویوکوکوس اوتیتیدیس یک باکتری سخت رشد، کند رشد و یکی از عوامل مهم ایجاد کننده عفونت گوش میانی است. شناسایی الویوکوکوس به روش کشت بسیار دشوار است در حالی کهPCR روش مناسب تری برای شناسایی این باکتری است.
کلید واژگان: الویوکوکوس اوتیتیدیس, اوتیت میانی, PCR, کشتBackground And AimIn Iran vaccination with Bacillus Calmette-Guérin (BCG) is performed on all newborns within the first days of life for prevention of tuberculosis. It is a live attenuated vaccine and produced from genetically different vaccine strains of Mycobacterium bovis. This vaccine is safe but local adverse reactions such as administration site abscess and lymphadenitis occur in some healthy children vaccinated with this vaccine as the most common side effect of BCG vaccination. Disseminated BCG infections are very rare in immunocompetent children but lymphadenitis is very common in Iran and other countries. It is indicated that the vaccine strain and it’s genetically variations are correlated with these side effects. Therefore, in this study we aimed to analyze the genetic characterizations of vaccine strains used in Iran.
Materials And MethodsThirty infants showing lymphadenitis induced by BCG vaccination were chosen for this study. After aspiration from the lesions, they were subjected to acid fast staining and culture on Lowenstein Jensen medium. Subsequently, DNA was extracted from samples using Phenol–chloroform method. The genus of the isolates was characterized by primer for 16sRNA gene. Then using RD1, RD14 and DU1 primers and in the next step RD9, RD4Deleted, RD4Present and RD1Deleted primers the species and strains of the isolates were characterized.
ResultsPerforming 16sRNA PCR, all 30 samples of acid fast bacilli were confirmed as Mycobacterium genus. Then using RD1, RD14, DU1, RD9, RD4 Deleted, RD4 Present and RD1 Deleted primers all isolates were detected as Mycobacterium bovis BCG strain Pasteur.
ConclusionIn this study all of the strains isolated from the patients were detected as Mycobacterium bovis BCG strain Pasteur. Therefore, the other possible factors causing BCG complications including BCG overdose, faulty intradermal technique, and disturbance of cellular immunity should be considered as the other risk factors of causing lesions.
Keywords: Mycobacterium bovis, BCG vaccine, Lymphadenitis, Tuberculosis, Infants -
زمینه و هدفروش MIRU-VNTR به یکی از فراگیرترین روش های استاندارد شده ژنوتایپینگ مایکوباکتریوم توبرکلوزیس کمپلکس تبدیل شده است. این مطالعه به منظور تعیین ساختار ژنتیکی مایکوباکتریوم توبرکلوزیس به روش MIRU-VNTR و نیز تعیین نقش اعضای دیگر مایکوباکتریوم توبرکلوزیس کمپلکس در ایجاد بیماری سل انجام گردید.روش بررسیاین مطالعه توصیفی روی 53 جدایه حاصل از کشت باکتریایی نمونه های خلط و لاواژ معدی بیماران مشکوک به بیماری سل انجام شد. به منظور تعیین هویت جدایه ها در ابتدا از آزمون های 16S rRNA و Rv Typing و در مرحله بعد از آزمون RD typing استفاده شد. سپس با به کارگیری 12 لوکوس شناخته شده MIRU-VNTR typing ژنوتایپ جدایه ها مشخص گردید.یافته هادر مجموع 44 تیپ ژنتیکی مورد شناسایی قرار گرفت. به طوری که 13 جدایه در 4 تیپ ژنتیکی به صورت مشترک یک ژنوتیپ را از خود نشان دادند و 40 تیپ ژنتیکی هر کدام فقط توسط یک جدایه نمایش داده شدند. در مقایسه میان لوکوس های 12 گانه MIRU-VNTR از نظر قدرت تفریق مشخص گردید MIRU-26 با 7 آلل قدرتمندترین توان افتراق میان جدایه ها را از خود نشان داده است. به طوری که Simpson’s diversity index در این لوکوس عدد 0.767 بود. به جز یک جدایه با هویت مایکوباکتریوم بوویس، 52 جدایه دیگر مایکوباکتریوم توبرکلوزیس بودند. هیچ یک از نمونه ها به سایر اعضای مایکوباکتریوم توبرکلوزیس کمپلکس آلوده نبودند. همچنین آلودگی همزمان نمونه ها به بیش از دو سویه مشاهده نشد.نتیجه گیریاگرچه 42 تیپ ژنتیکی MIRU-VNTR در میان 53 جدایه تحت مطالعه شناسایی شد؛ اما دستکم در 19 مورد تفاوت میان این تیپ ها، فقط در مورد یک لوکوس از 12 لوکوس به کاررفته مشاهده گردید. بدین ترتیب در مجموع جمعیت ژنتیکی نسبتا همگنی از جدایه ها مشاهده گردید. اگرچه شناسایی 13 جدایه اپیدمیک در قالب 4 تیپ ژنتیکی می تواند به عنوان نشانه انتقال بیماری در میان مبتلایان باشد؛ اما در مجموع انتقال بیماری در بیماران تحت بررسی چندان گسترده به نظر نمی رسد.
کلید واژگان: مایکوباکتریوم توبرکلوزیس کمپلکس, مایکوباکتریوم بوویس, RD typing, Rv typing, MIRU, VNTR genotypingBackground And ObjectiveMIRU-VNTR typing is currently one of the most frequently-used standardized genotyping systems in molecular epidemiology of tuberculosis in the world. This sudy was done to determine the Mycobacterium tuberculosis genotyping by MIRU-VNTR method.MethodsThis descriptive study was done on sputum, gastric lavage clinical specimens of 53 tuberculosis suspected patients. Fifty-three isolates were identified by 16S rRNA and Rv-typing followed by RD typing. They were then subjected to a 12-locus (ETRA, ETRB, ETRC, ETRD, ETRE and ETRF, MIRU-10, MIRU-26, MIRU-39, MIRU-30 plus QUB-11b) MIRU-VNTR typing system.ResultsIn MIRU-VNTR typing, forty-four types were identified with 13 isolates classified in 4 clustered and the remaining 40 isolates representing 40 orphan patterns. In comparative analysis of MIRU-VNTR loci, MIRU-26 with 7 alleles displayed the highest diversity level (Simpson’s diversity index = 0.767. Out of the 53 isolates, only one was identified as Mycobacterium bovis. All the remaining isolates were characterized as Mycobacterium tuberculosis. None of the samples was affected to Mycobacterium complex strain. No evidence of either double or co-infection of the patients with more than one species/strain was detected.ConclusionWhile the genomic diversity observed by MIRU-VNTR typing sounds extensive, the population genomic structure on the whole however, seems to be homogenous. Recent transmission between studied patients does not appear to be a frequent event as only 13 isolates representing 4 MIRU-VNTR types, were assumingly epidemic.Keywords: Mycobacterium tuberculosis complex, Mycobacterium bovis, RD typing, Rv, typing, MIRU, VNTR genotype -
مقدمه و اهدافسل یکی از مهم ترین بیماری های عفونی و زئونوز است که طیف وسیعی از موجودات زنده به ابتلا به آن حساس هستند. پژوهش حاضر با هدف بررسی عوامل موثر بر آلودگی گاوداری های تحت پوشش آزمون غربالگری توبرکولین به سل گاوی انجام شد.روش کاراین پژوهش، یک مطالعه مورد- شاهدی است که در سطح گله های گاو شیری انجام شد. گاوداری های مورد (62 فارم) از میان گاوداری های شیری تحت پوشش استان تهران و 7 استان هم جوار انتخاب شد که پس از نمونه برداری، مایکوباکتریوم بویس از آن ها در جدا گردید. گاوداری های شاهد (62 فارم) نیز از همین استان ها و بر اساس منفی بودن نتیجه آزمون توبرکولین انجام پذیرفت. گاوداری های شاهد بر اساس ظرفیت و فاصله با گاوداری های مورد، به صورت فردی هم سان شدند. تجزیه و تحلیل داده ها با آزمون رگرسیون لجستیک شرطی چند متغیره و نرم افزارStata انجام شد.نتایجاز نظر روابط بین متغیرهای مستقل تحت مطالعه، مدیریت مناسب کود (OR=0.12; 95% CI: 0.03-0.49)، شعله دهی منظم بهاربندها (OR= 0.21; 95% CI: 0.04-0.92)، حصارکشی منظم اطراف دامداری (OR= 0.17; 95% CI: 0.03-0.81)، سبب کاهش خطر آلودگی و وجود موش (OR= 4.90; 95% CI: 1.04-23.01) سبب افزایش خطر آلودگی به سل گاوی می شود.نتیجه گیریتوجه جدی تر به مبارزه با جوندگان محسوس است. افزایش همه جانبه برنامه ی غربالگری توصیه می شود. شاید زمان آن فرا رسیده باشد تا در سیاست های موجود کنترلی بازنگری صورت پذیرد و سیاست های مناسب تری اتخاذ گردد.
کلید واژگان: مایکوباکتریوم بویس, سل, گاو, مطالعه مورد, شاهدی هم سان, رگرسیون لجستیک شرطیBackground and ObjectivesBovine Tuberculosis (BTB) is one of the most important zoonoses. Mycobacterium bovis is the responsible agent of BTB in the cattle. The current study was conducted to investigate the determination factors of BTB in dairy farms covered by the tuberculin screening test.MethodsA herd level case- control study was carried out in 124 (62 cases & 62 controls) dairy farms in the provinces of Tehran, Alborz, Hamedan, Isfahan, Qazvin, Qom, Mazandaran and Semnan. The control farms were individually matched with case farms by farm capacity and distance. Statistical analyses were done by Stata 11.2 using conditional logistic regression.ResultsProper management of manure (OR=0.12; 95% CI: 0.03-0.49), regular flaming of stalls (OR= 0.21; 95% CI: 0.04-0.92) and complete fencing around the farm (OR= 0.17; 95% CI: 0.03-0.81) decreased while the presence of rodents (rat) (OR= 4.90; 95% CI: 1.04-23.01) increased the risk of infection. The interaction among these variables was not statistically significantConclusionAccording to the results, there is an essential need to pay more attention to rodent control in farms.Keywords: Mycobacterium bovis, Tuberculosis, Matched Case, control study, Conditional logistic regression -
BackgroundBCG (Bacille Calmette-Guerin) vaccine is an attenuated live vaccine administered to prevent Tuberculosis. Disseminated infection due to BCG is a rare life threatening complication of this vaccine, especially in immunocompromised patients..ObjectivesThe current study evaluated the sensitivity pattern of the BCG strain supplied by the Institute de Pasteur, Iran against anti-mycobacterial drugs used to treat disseminated infection caused by the vaccine..Patients and Materials: Bacille Calmette-Guerin strain ATCC 1173 P was purchased from the Pasteur Institute in Tehran and three samples of Mycobacterium bovis isolated from subaxillary adenitis of three patients were tested for susceptibility to major anti-mycobacterial drugs by MODS (microscopic observation drug sensitivity) method..ResultsAll samples of BCG strain were resistant to most of the anti-mycobacterial drugs used separately except for concentrations of isoniazid 1 µg/mL, ethambutol 3 µg/mL, ciprofloxacin 4 µg/mL and clarithromycin 2 µg/mL. Addition of pyrazinamide (PZA) 25 µg/mL, the susceptibility pattern did not change to any drug, but increasing PZA concentration to 50 µg/ml and combining it with ethionamide 2 µg/mL, isoniazid 0.2 µg/mL, streptomycin 2 µg/mL, rifampin 0.5 µg/mL or ciprofloxacin 0.2 µg/mL, made the Mycobacterium strains susceptible..ConclusionsThe current study findings revealed that Mycobacterium bovis resistance to pyrazinamide is resolved by increasing the concentration of the drug and combining PZA with other anti-mycobacterial agents..Keywords: Mycobacterium bovis, Pyrazinamide, Resistance
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BackgroundMycobacterium bovis is the causative agent of bovine tuberculosis and belongs to the Mycobacterium tuberculosis complex. M. bovis usually carries only one or a few copies of the insertion sequence IS6110 in its genome. The aim of this study was evaluation of copy number of IS6110 in M. bovis isolates by restriction fragment length polymorphism (RFLP) method.Materials And MethodsIn this experimental study, 25 lymph node specimens of tuberculin-positive cattle were collected and cultured by standard methods, afterward genomic DNA was extracted by chloroform-isoamyl alcohol. Genetic studies were conducted by Pvull and DNA hybridization with IS6110.ResultsTwo isolates displayed more than of 4 copies of IS6110 by RFLP (IS6110-RFLP) method.ConclusionThe results of this study are unique and specific in Iran but reported in the world rarely. Therefore the new strains of M. bovis imported to Iran from other countries of the world.Keywords: Mycobacterium bovis, Restriction fragment length polymorphism, Tuberclusis
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