Name: system for efficacy and cytotoxicity screening of inhibitors targeting intracellular mycobacterium tuberculosis
Uploaded: 2017-04-05T14:30:00
Description: Watch this Scientific Journal Video about System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis at JoVE.com

This work was supported by BC Lung Association and Mitacs.

1. سلالة بكتيرية والنمو المتوسطة <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> جعل سكر العنب الزلال وحل الأسهم الملح (ADS) من خلال الانحلال 25 غرام من الزلال المصل البقري، 10.0 غرام من سكر العنب، و4.05 غرام من كلوريد الصوديوم في 460 مل من الماء منزوع الأيونات. تصفية تعقيم AD...

<ol>
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S., Alber, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+tyrosine+phosphatase+PtpA+is+not+required+for+Mycobacterium+tuberculosis+growth+in+mice.">Protein tyrosine phosphatase PtpA is not required for Mycobacterium tuberculosis growth in mice.</a> FEMS Microbiol Lett. 287, 181-184 (2008).</li><li>Wong, D., Bach, H., Sun, J., Hmama, Z., Av-Gay, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mycobacterium+tuberculosis+protein+tyrosine+phosphatase+(PtpA)+excludes+host+vacuolar-H+-ATPase+to+inhibit+phagosome+acidification.">Mycobacterium tuberculosis protein tyrosine phosphatase (PtpA) excludes host vacuolar-H+-ATPase to inhibit phagosome acidification.</a> Proc Natl Acad Sci U S A. 108, 19371-19376 (2011).</li><li>Bach, H., Papavinasasundaram, K. 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وكان الهدف من هذه الدراسة هو إيجاد طريقة HTS بسيطة وفعالة من حيث التكلفة باستخدام نموذج العدوى داخل الخلايا البشرية لM. السل. والسل هو مرض يصيب الإنسان تتميز إصابة الضامة السنخية التي كتبها M. السل. بسبب قضايا السلامة الأحيائية، وقد استخدم البحوث التي تنطوي عل...

المتفطرة السلية، العامل المسبب لمرض السل (TB)، هو أحد الأسباب الرئيسية للمراضة والوفيات في جميع أنحاء العالم. TB الحساس للأدوية هو مرض قابل للعلاج يتطلب مضادات حيوية متعددة لمدة 6 أشهر. على الرغم من كونه مرض قابل للعلاج، وقدرت نسبة الوفيات بمرض السل أن يكون 1.5 مليون في عام 2015 1. في السنوات ال 10 الماضية، كانت هناك زيادة المخاوف من انتشار M. السل المقاوم للأدوية. يتم تعريف للأدوية المتعددة مقاومة السل (MDR-TB) كما TB التي هي مقاومة على الأقل أيزونيازيد (INH) وريفامبيسين (RMP)، ومعظم سلالات السل المقاوم للأدوية المتعددة هي أيضا مقاومة لتحديد الأدوية TB الخط الثاني، مما يحد من خيارات العلاج . آثار مقاومة المخدرات تخلق تحديا أكبر لعلاج المرضى الذين يعانون شارك في المصابين بفيروس نقص المناعة البشرية (HIV). 400000 المرضى في جميع أنحاء العالم توفي من مرض السل المرتبطة بفيروس نقص المناعة البشرية في عام 2015 1. مخيبة للآمال، والولايات المتحدة الأمريكية للغذاء والدواء Administوقد وافقت التموينية واحد فقط المخدرات TB جديدة ضد MDR-TB، bedaquiline، في السنوات ال 40 الماضية 2. هناك حاجة ماسة إلى التقدم في إيجاد علاجات أفضل وأقصر TB من أجل البقاء قدما في مكافحة السل وMDR-TB. تقليديا، يتم تنفيذ الشاشات المخدرات TB تحت في المختبر ظروف النمو في متوسط النمو، حيث تتم إضافة مركبات للثقافة متنامية ويتم تحديد فعاليتها في القضاء على الكائنات الحية الدقيقة عن طريق عد مستعمرة (كفو) في وسط صلب. كما تعول CFU هي كثيفة العمالة، تستغرق وقتا طويلا، ومكلفة، وقد وضعت وسائل غير مباشرة مختلفة للتخفيف من حدة هذه المشكلة. وتشمل هذه الأساليب في الامار الأزرق جدوى فحص 3، وتحديد مضان 4 من البروتين الأخضر الفلورسنت (GFP) أو التلألؤ 5 من البكتيريا، معربا عن وسيفيراز، وتقدير مجموع أدينوسين ثلاثي الفوسفات (ATP) 6 و 7. يتميز TB نموذجي من عدوى السل M. في الرئة، حيث تتواجد البكتيريا وتتكاثر داخل جسم بلعمي الضامة السنخية 8. الشاشة المظهرية بسيطة في مرق قد تتناسب مع مسببات الأمراض خارج الخلية. ومع ذلك، في المنظور التاريخي، وأصيب المركبات ضد M. السل التي تم تحديدها باستخدام هذه الطريقة غالبا ما تفشل في الارتقاء إلى مستوى التوقعات خلال خطوات التحقق من صحة المصب في نماذج العدوى. نقترح أن المخدرات TB يتم تنفيذ الأفضل في نموذج عدوى الخلية المضيفة داخل الخلايا. ومع ذلك، ونماذج الخلايا تمتلك العديد من الحواجز التكنولوجية والبيولوجية لفحص (HTS) تطوير الإنتاجية العالية. وهناك عقبة كبيرة هي تعقيد عملية العدوى، والتي تجسدت خطوات عديدة وإزالة متطورة من البكتيريا خارج الخلية التي كتبها في الفترات الفاصلة بين الغسيل. وهناك عقبة رئيسية الثانية هي المرة إعادة طويلةيتطلبه، كما كشف عن النمو، ويتم عادة CFU تعول على لوحات ثقافة، هي العملية التي تستغرق أكثر من 3 أسابيع لإكمال. تم توفير حل واحد ليحل محل التهم CFU بواسطة المجهر الفلورسنت الآلي في تركيبة مع البكتيريا الفلورسنت. ومع ذلك، هذا الحل يتطلب الاستثمار في المعدات الأولي الذي هو خارج عن متناول العديد من مختبرات البحوث. ومن شأن ومنخفضة التكلفة بسيطة، والأمراض ذات الصلة طريقة HTS يعزز إلى حد كبير عملية اكتشاف المخدرات. في هذه الدراسة، ونحن التقرير نظام HTS جديد، وحدات التي تهدف إلى توفير السريع، وتدرجية عالية، بعد اقتصادي، فحص مناسبة لتحديد النشاط من المركبات ضد الخلايا السل M.. ويتكون هذا النظام من ثلاث وحدات: (ط) داخل الخلايا، (ب) السمية الخلوية، و (iii) فحوصات في المرق. توفر النتيجة النهائية مجتمعة وصفا شاملا للخصائص مركب، مع معلومات إضافية عن الوضع المحتمل للعمل. هذا الشورينظام reening استخدمت في العديد من المشاريع مع المكتبات مجمع المختلفة التي تستهدف طريقة العمل، بما في ذلك تحليل التآزر المخدرات 9، وتحفيز الالتهام الذاتي 10، وتثبيط M. السل -secreted عامل الضراوة (غير منشورة). كما تم دراسة المركبات من وضع غير معروف من عمل 11. اعتمد نسخة معدلة من هذا الأسلوب أيضا شريك صناعي لدينا كأسلوب غربلة أولية لتحديد مركبات جديدة ضد الخلايا السل M. 11.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>RPMI 1640</td>
			<td>Sigma-Aldrich</td>
			<td>R5886</td>
			<td></td>
		</tr>
		<tr>
			<td>L-glutamine</td>
			<td>Sigma-Aldrich</td>
			<td>G7513</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum (FBS)</td>
			<td>Thermo Fisher Scientific</td>
			<td>12483020</td>
			<td></td>
		</tr>
		<tr>
			<td>Middlebrook 7H9</td>
			<td>Becton, Dickinson and Company</td>
			<td>271210</td>
			<td></td>
		</tr>
		<tr>
			<td>Tween80</td>
			<td>Fisher Scientific</td>
			<td>T164</td>
			<td></td>
		</tr>
		<tr>
			<td>Albumin, Bovine pH7</td>
			<td>Affymetrix</td>
			<td>10857</td>
			<td></td>
		</tr>
		<tr>
			<td>Dextrose</td>
			<td>Fisher Scientific</td>
			<td>BP350</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride</td>
			<td>Fisher Scientific</td>
			<td>BP358</td>
			<td></td>
		</tr>
		<tr>
			<td>kanamycin sulfate</td>
			<td>Fisher Scientific</td>
			<td>BP906</td>
			<td></td>
		</tr>
		<tr>
			<td>PMA</td>
			<td>Sigma-Aldrich</td>
			<td>P8139</td>
			<td></td>
		</tr>
		<tr>
			<td>MTT</td>
			<td>Sigma-Aldrich</td>
			<td>M2128</td>
			<td></td>
		</tr>
		<tr>
			<td>N,N-Dimethylformamide (DMF)</td>
			<td>Fisher Scientific</td>
			<td>D131</td>
			<td></td>
		</tr>
		<tr>
			<td>1M Hydrocholoric acid (HCl)</td>
			<td>Fisher Scientific</td>
			<td>351279212</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetic acid</td>
			<td>Fisher Scientific</td>
			<td>351269</td>
			<td></td>
		</tr>
		<tr>
			<td>SDS</td>
			<td>Fisher Scientific</td>
			<td>BP166</td>
			<td></td>
		</tr>
		<tr>
			<td>Resazurin</td>
			<td>Alfa Aesar</td>
			<td>B21187</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>D5879</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycerol</td>
			<td>Fisher Scientific</td>
			<td>BP229</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>THP-1</td>
			<td>American Type Culture Collection</td>
			<td>TIB-202</td>
			<td></td>
		</tr>
		<tr>
			<td>M. tuberculosis H37Rv</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>96-well flat bottom white plate</td>
			<td>Corning</td>
			<td>3917</td>
			<td></td>
		</tr>
		<tr>
			<td>95-well flat bottom clear plate</td>
			<td>Corning</td>
			<td>3595</td>
			<td></td>
		</tr>
		<tr>
			<td>Transparent plate sealer</td>
			<td>Thermo Fisher Scientific</td>
			<td>AB-0580</td>
			<td></td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer</td>
			<td>Thermo Fisher Scientific</td>
			<td>Biomate 3</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate spectrophotometer</td>
			<td>Biotek</td>
			<td>Epoch</td>
			<td></td>
		</tr>
		<tr>
			<td>luminometer</td>
			<td>Applied Biosystems</td>
			<td>Tropix TR717</td>
			<td></td>
		</tr>
	</tbody>
</table>

system for efficacy and cytotoxicity screening of inhibitors targeting intracellular mycobacterium tuberculosis

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a leading cause of morbidity and mortality worldwide. With the increased spread of multi drug-resistant TB (MDR-TB), there is a real urgency to develop new therapeutic strategies against M. tuberculosis infections. Traditionally, compounds are evaluated based on their antibacterial activity under in vitro growth conditions in broth; however, results are often misleading for intracellular pathogens like M. tuberculosis since in-broth phenotypic screening conditions are significantly different from the actual disease conditions within the human body. Screening for inhibitors that work inside macrophages has been traditionally difficult due to the complexity, variability in infection, and slow replication rate of M. tuberculosis. In this study, we report a new approach to rapidly assess the effectiveness of compounds on the viability of M. tuberculosis in a macrophage infection model. Using a combination of a cytotoxicity assay and an in-broth M. tuberculosis viability assay, we were able to create a screening system that generates a comprehensive analysis of compounds of interest. This system is capable of producing quantitative data at a low cost that is within reach of most labs and yet is highly scalable to fit large industrial settings.

الإنتاجية العالية فحص الخلايا باستخدام M. السل التعبير عن الجينات وسيفيراز الشكل 2A والجدول 1 يحتوي على البيانات الخام التي تم جمعها من قبل luminometer، أعرب في وحدات الانارة ال?...

Watch this Scientific Journal Video about System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis at JoVE.com

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

لقد قمنا بتطوير نظام الفحص الإنتاجية العالية وحدات لاكتشاف مركبات جديدة ضد السل المتفطرة، تستهدف داخل وفي مرق تزايد البكتيريا وكذلك السمية الخلوية ضد الخلية المضيفة الثدييات.

نظام لفعالية والسمسة فحص مثبطات استهداف بين الخلايا<em&gt; المتفطرة السلية</em

Mycobacterium tuberculosis, the causative agent of tuberculosis (TB), is a leading cause of morbidity and mortality worldwide. With the increased spread ...

The overall goal of this protocol is to provide multidimensional assessment of how compounds affect M.tuberculosis survival inside human macrophages. This method is centered on a Luciferase-based efficacy assay. When combined with cytotoxicity an MIC assay can provide useful information into modification of HIT compounds in TB drug development.The main advantage of our method is that it is time and labor saving. It is replacing a CFU colony forming unit assay with a Luciferase based assay. Generally individuals new to this technique will struggle with consistency with manual pipetting instead of automation.A consistent pipetting technique is crucial. Grow THP1 cells in RPMI in complete medium. While maintaining a cell density of 0.2 to one million cells per milliliter of medium between passages.Use a spectrophotometer to measure the OD600 of an actively growing bacterial suspension. Then calculate the bacterial density using the conversion factor of 0.1 OD600 equals three times 10 to the seventh bacteria per milliliter. Pipette out sufficient bacteria for an MOI of 10 to one into a new centrifuge tube.Then pellet the bacteria at 3, 000 times G for 10 minutes. To opsonize the bacteria, add 50 microliters of human serum to 450 microliters of RPMI 1640 and use it to resuspend the bacterial pellet at a density of one times 10 to the eighth bacteria per 500 microliters of medium. Allow the mixture to incubate at 37 degrees Celsius for 30 minutes.Next, using a hemocytometer and an inverted microscope, determine the THP1 cell culture density. Then, in sterile centrifuge tubes, pellet the cells at 100 times G in 37 degrees Celsius for 10 minutes. Aspirate the supernatant and resuspend the cells in RPMI in complete medium at a density of one million cells per milliliter.Add PMA to a final concentration of 40 nanograms per milliliter. This is differentiation mix. Combined opsonized M.tuberculosis with THP1 differentiation mix at an MOI of 10 to one.And aliquot the final mix at 100 microliters per well in a 96 well white flat bottom plate. Then allow the differentiation and infection to incubate at 37 degrees Celsius in a humidified incubator containing 5%carbon dioxide overnight. The following day, use 100 microliters of RPMI to wash the wells twice.Then add compounds diluted to the desired concentrations and RPMI in complete medium. Incubate the plates for three days. When using multichannel pipettes, remember to practice slow and steady plunger action and always land the pipette tips in the same spot inside each well to minimize disturbance to the THP1 cells.Following the incubation, aspirate the medium from the wells and add 50 microliters of luciferase assay reagent to each well. Then use transparent adhesive plate sealers to seal the plates. Incubate the plates at 22 degrees Celsius for five minutes, then use a luminometer to obtain a readout at one second per well.To differentiate THP1 cells, after preparing differentiation mix as demonstrated earlier in this video, dispense 100 microliters of the differentiation mix into each well of a clear, 96 well plate of cells. Incubate the cells at 37 degrees Celsius in a humidified incubator with 5%carbon dioxide overnight. The following day, aspirate the medium from the wells and use RPMI 1640 to wash them twice.Add 100 microliters of compounds diluted in RPMI incomplete to the wells. Then, incubate the cells for three days. Dissolve 0.5 grams of MTT in 100 milliliters of PBS to make a stock solution of five milligrams per milliliter.Then filter sterilize the solution. Two and a half hours before the end of the three day incubation period, add 25 microliters of MTT solution to each well of cells and complete the incubation period. Prepare 50%NN dimethylformamide or DMF by mixing 250 milliliters of DMF with 250 milliliters of deionized water.Prepare MTT extraction buffer as follows. Lay 100 grams of SDS in a 500 milliliter bottle and add 300 milliliters of 50%DMF. Apply low heat to allow the SDS to dissolve.Add 10 milliliters of pure acetic acid and 12.5 milliliters of one molar hydrochloric acid. Then, use 50%DMF to fill up the bottle to the 500 milliliter mark. At the end of the treatment period, add 100 microliters of extraction buffer to each well.Incubate the mixture at 37 degrees Celsius in a humidified incubator containing 5%carbon dioxide overnight. The following day, read the absorbance at 570 nanometers. To carry out a resazurin assay, grow M.tuberculosis in 7H9 ADST to midlock phase.Dilute the culture with 7H9 ADST to an OD600 of 0.01. Use 7H9 ADST to dilute the compounds to two times the testing concentrations. And aliquot 100 microliters of each diluted compound into each well of a clear 96 well plate.Transfer 100 microliters of the diluted bacterial suspension into each well. Incubate the plates at 37 degrees Celsius in a humidified incubator for five days. Dissolve 10 milligrams of resazurin in 100 milliliters of deionized water.And filter sterilize the solution. Add 30 microliters of resazurin solution to the wells. Then incubate the cells for 48 hours.Bacterial growth is indicated by a color conversion from blue to pink. Carry out quantitative analysis according to the text protocol. This scatterplot shows the raw luminescence data measured in relative luminescence units for the treatment of various concentrations of rifampicin on M.tuberculosis and THP1 cells.In this plot, the percent reduction and luminescence in treated wells compared to untreated wells is shown. Rifampicin is capable of reducing 99.9%of the relative light units, or RLUs from intracellular M.tuberculosis at a concentration of 0.1 micrograms per milliliter. Similar to previously published results determined from CFU.In this figure, the cytotoxicity caused by increasing concentrations of G1-1H used in the MTT assay is illustrated. Concentrations of 10 and three micromolar are clearly below the IC 50 cutoff. This figure shows the effects of various concentrations of the antibiotic apramycin on resazurin conversion by M.tuberculosis.The in-broth MIC90 was between 2.5 and five micrograms per milliliter. This is slightly higher than the previously published value of 1.5 micrograms per milliliter. But it is still well within the acceptable range.As demonstrated here, evaporation becomes significant for all experiments with prolonged incubation times. So care must be taken to keep incubators well humidified to avoid inconsistencies. Once mastered, this technique can provide reliable intracellular data in as little as four days.While attempting this procedure it is important to remember to be gentle with THP1 cells, and be consistent with pipetting if attempting without automation. Following this procedure, other methods like high content screening can be used to determine if hit compounds are bacteriostatic or bactericidal. After its development, this protocol paved the way for researchers in the field of tuberculosis drug discovery to rapidly assess compound efficacy against MTB inside human macrophages.Don't forget that working with microbacterium tuberculosis is dangerous. Use precautions and proper equipment when handling this microorganism.

Research

JoVE Journal

Immunology and Infection

System for Efficacy and Cytotoxicity Screening of Inhibitors Targeting Intracellular Mycobacterium tuberculosis

Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies

Sample Preparation of Mycobacterium tuberculosis Extracts for Nuclear Magnetic Resonance Metabolomic Studies

تحضير العينة من<em&gt; المتفطرة السلية</em&gt; خلاصات لدراسات النووية المغناطيسي الرنين Metabolomic

Mycobacterium tuberculosis is a major cause of mortality in human beings on a global scale. The emergence of both multi- (MDR) and extensively-(XDR) ...

Growth of Mycobacterium tuberculosis Biofilms

Growth of Mycobacterium tuberculosis Biofilms

نمو المتفطرة السلية الأغشية الحيوية

Mycobacterium tuberculosis, the etiologic agent of human tuberculosis, has an extraordinary ability to survive against environmental stresses including ...

An Experimental Model to Study Tuberculosis-Malaria Coinfection upon Natural Transmission of Mycobacterium tuberculosis and Plasmodium berghei

An Experimental Model to Study Tuberculosis-Malaria Coinfection upon Natural Transmission of Mycobacterium tuberculosis and Plasmodium berghei

نموذج تجريبي لدراسة الملاريا والسل عدوى مرافقة على انتقال الطبيعية السل المتفطرة و المتصورة berghei</i

Coinfections naturally occur due to the geographic overlap of distinct types of pathogenic organisms. Concurrent infections most likely modulate the ...

High-throughput Screening for Broad-spectrum Chemical Inhibitors of RNA Viruses

الإنتاجية العالية للفحص واسع الطيف الكيميائية مثبطات الحمض النووي الريبي الفيروسات

RNA viruses are responsible for major human diseases such as flu, bronchitis, dengue, Hepatitis C or measles. They also represent an emerging threat ...

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

Demonstrating a Multi-drug Resistant Mycobacterium tuberculosis Amplification Microarray

مما يدل على متعدد المخدرات مقاومة<em&gt; السل المتفطرة</em&gt; التضخيم ميكروأري

Simplifying microarray workflow is a necessary first step for creating MDR-TB microarray-based diagnostics that can be routinely used in lower-resource ...

Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors

الفحص السريع للفيروس نقص المناعة البشرية والناسخ العكسي مثبطات Integrase

Although a number of anti HIV drugs have been approved, there are still problems with toxicity and drug resistance. This demonstrates a need to identify ...

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

نموذج الأنسجة 3D الرئة الإنسان للدراسات وظيفية على<em&gt; المتفطرة السلية</em&gt; العدوى

Tuberculosis (TB) still holds a major threat to the health of people worldwide, and there is a need for cost-efficient but reliable models to help us ...

A High-throughput Compatible Assay to Evaluate Drug Efficacy against Macrophage Passaged Mycobacterium tuberculosis

A High-throughput Compatible Assay to Evaluate Drug Efficacy against Macrophage Passaged Mycobacterium tuberculosis

وإنتاجية عالية الفحص المدعم لتقييم فعالية الدواء ضد بلعم Passaged<em&gt; المتفطرة السلية</em

The early drug development process for anti-tuberculosis drugs is hindered by the inefficient translation of compounds with in vitro activity to ...

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

Analysis of 18FDG PET/CT Imaging as a Tool for Studying Mycobacterium tuberculosis Infection and Treatment in Non-human Primates

تحليل 18"فدج التصوير" PET/CT كأداة لدراسة بكتريا السل العدوى والعلاج في الإنسان عدم الرئيسات

Mycobacterium tuberculosis remains the number one infectious agent in the world today. With the emergence of antibiotic resistant strains, new clinically ...

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

Imaging Mycobacterium tuberculosis in Mice with Reporter Enzyme Fluorescence

تصوير بكتريا السل في الفئران مع "مراسل إنزيم الأسفار"

Reporter enzyme fluorescence (REF) utilizes substrates that are specific for enzymes present in target organisms of interest for imaging or detection by ...