scholarly journals Novel T7 Phage Display Library Detects Classifiers for Active Mycobacterium Tuberculosis Infection

Viruses ◽  
2018 ◽  
Vol 10 (7) ◽  
pp. 375 ◽  
Author(s):  
Harvinder Talwar ◽  
Samer Hanoudi ◽  
Sorin Draghici ◽  
Lobelia Samavati
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Microarray Platform ◽  
Phage Display Library ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Operating Characteristics ◽  
Mycobacterium Tuberculosis Infection ◽  
Bodily Fluids ◽  
T7 Phage Display ◽  
T7 Phage

Tuberculosis (TB) is caused by Mycobacterium tuberculosis (MTB) and transmitted through inhalation of aerosolized droplets. Eighty-five percent of new TB cases occur in resource-limited countries in Asia and Africa and fewer than 40% of TB cases are diagnosed due to the lack of accurate and easy-to-use diagnostic assays. Currently, diagnosis relies on the demonstration of the bacterium in clinical specimens by serial sputum smear microscopy and culture. These methods lack sensitivity, are time consuming, expensive, and require trained personnel. An alternative approach is to develop an efficient immunoassay to detect antibodies reactive to MTB antigens in bodily fluids, such as serum. Sarcoidosis and TB have clinical and pathological similarities and sarcoidosis tissue has yielded MTB components. Using sarcoidosis tissue, we developed a T7 phage cDNA library and constructed a microarray platform. We immunoscreened our microarray platform with sera from healthy (n = 45), smear positive TB (n = 24), and sarcoidosis (n = 107) subjects. Using a student t-test, we identified 192 clones significantly differentially expressed between the three groups at a False Discovery Rate (FDR) <0.01. Among those clones, we selected the top ten most significant clones and validated them on independent test set. The area under receiver operating characteristics (ROC) for the top 10 significant clones was 1 with a sensitivity of 1 and a specificity of 1. Sequence analyses of informative phage inserts recognized as antigens by active TB sera may identify immunogenic antigens that could be used to develop therapeutic or prophylactic vaccines, as well as identify molecular targets for therapy.

scholarly journals Detection of Cystic Fibrosis Serological Biomarkers Using a T7 Phage Display Library

2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Harvinder Talwar ◽  
Samer Najeeb Hanoudi ◽  
Andreea Geamanu ◽  
Dana Kissner ◽  
Sorin Draghici ◽  
...  
Keyword(s):  
Cystic Fibrosis ◽  
Phage Display ◽  
Phage Display Library ◽  
T7 Phage Display ◽  
Serological Biomarkers ◽  
T7 Phage

Production, characterization and in-vitro applications of single-domain antibody against thyroglobulin selected from novel T7 phage display library

2021 ◽  
Vol 492 ◽  
pp. 112990
Author(s):  
Jothivel Kumarasamy ◽  
Samar Kumar Ghorui ◽  
Chandrakala Gholve ◽  
Bharti Jain ◽  
Yogesh Dhekale ◽  
...  
Keyword(s):  
Phage Display ◽  
Phage Display Library ◽  
Single Domain ◽  
Single Domain Antibody ◽  
Domain Antibody ◽  
T7 Phage Display ◽  
T7 Phage

scholarly journals Characterization of Drug-Resistant Lipid-Dependent Differentially Detectable Mycobacterium tuberculosis

2021 ◽  
Vol 10 (15) ◽  
pp. 3249
Author(s):  
Annelies W. Mesman ◽  
Seung-Hun Baek ◽  
Chuan-Chin Huang ◽  
Young-Mi Kim ◽  
Sang-Nae Cho ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Culture Media ◽  
Multidrug Resistant ◽  
Culture Conditions ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Solid Culture ◽  
Lowenstein Jensen ◽  
Genetic Mechanisms ◽  
Lipid Resuscitation

An estimated 15–20% of patients who are treated for pulmonary tuberculosis (TB) are culture-negative at the time of diagnosis. Recent work has focused on the existence of differentially detectable Mycobacterium tuberculosis (Mtb) bacilli that do not grow under routine solid culture conditions without the addition of supplementary stimuli. We identified a cohort of TB patients in Lima, Peru, in whom acid-fast bacilli could be detected by sputum smear microscopy, but from whom Mtb could not be grown in standard solid culture media. When we attempted to re-grow Mtb from the frozen sputum samples of these patients, we found that 10 out of 15 could be grown in a glycerol-poor/lipid-rich medium. These fell into the following two groups: a subset that could be regrown in glycerol after “lipid-resuscitation”, and a group that displayed a heritable glycerol-sensitive phenotype that were unable to grow in the presence of this carbon source. Notably, all of the glycerol-sensitive strains were found to be multidrug resistant. Although whole-genome sequencing of the lipid-resuscitated strains identified 20 unique mutations compared to closely related strains, no single genetic lesion could be associated with this phenotype. In summary, we found that lipid-based media effectively fostered the growth of Mtb from a series of sputum smear-positive samples that were not culturable in glycerol-based Lowenstein–Jensen or 7H9 media, which is consistent with Mtb’s known preference for non-glycolytic sources during infection. Analysis of the recovered strains demonstrated that both genetic and non-genetic mechanisms contribute to the observed differential capturability, and suggested that this phenotype may be associated with drug resistance.

scholarly journals Pouched Rats’ Detection of Tuberculosis in Human Sputum: Comparison to Culturing and Polymerase Chain Reaction

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Amanda Mahoney ◽  
Bart J. Weetjens ◽  
Christophe Cox ◽  
Negussie Beyene ◽  
Klaus Reither ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Mycobacterium Tuberculosis ◽  
Sputum Smear ◽  
Smear Microscopy ◽  
Chain Reaction ◽  
Poor Countries ◽  
Valuable Adjunct ◽  
Polymerase Chain ◽  
The Individual ◽  
Pouched Rats

Setting. Tanzania.Objective. To compare microscopy as conducted in direct observation of treatment, short course centers to pouched rats as detectors ofMycobacterium tuberculosis.Design. Ten pouched rats were trained to detect tuberculosis in sputum using operant conditioning techniques. The rats evaluated 910 samples previously evaluated by smear microscopy. All samples were also evaluated through culturing and multiplex polymerase chain reaction was performed on culture growths to classify the bacteria.Results. The patientwise sensitivity of microscopy was 58.0%, and the patient-wise specificity was 97.3%. Used as a group of 10 with a cutoff (defined as the number of rat indications to classify a sample as positive forMycobacterium tuberculosis) of 1, the rats increased new case detection by 46.8% relative to microscopy alone. The average samplewise sensitivity of the individual rats was 68.4% (range 61.1–73.8%), and the mean specificity was 87.3% (range 84.7–90.3%).Conclusion. These results suggest that pouched rats are a valuable adjunct to, and may be a viable substitute for, sputum smear microscopy as a tuberculosis diagnostic in resource-poor countries.

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