Selection of Nucleic Acid Aptamers Specific for Mycobacterium tuberculosis

Mozioğlu, Erkan; Gokmen, Ozgur; Tamerler, Candan; Kocagoz, Zühtü; Akgöz, Müslüm

doi:10.1007/s12010-015-1913-7

Cited by 37 publications

(17 citation statements)

References 54 publications

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“…The SELEX (systematic evolution of ligands by exponential enrichment) technique was generated by using reiterative in vitro selection for high-a nity and -speci city oligonucleotide ligands (aptamers) against almost any kind of molecules that are of biological or therapeutic interest [35]. Aptamers provide a powerful tool not only used for identi cation of novel diagnostic markers but also for interference with the duplication of the pathogenic agents, and thus have been widely used as biorecognition probes against human pathogens and/or their toxins [36][37][38].…”

Section: Discussionmentioning

confidence: 99%

SELEX-Based Direct Enzyme-Linked Aptamer Assay(DELAA) for Diagnosis of Toxoplasmosis by Detection of SAG1 Antigen in Sera of Mice and Humans

Shen

Cui

Cudjoe

et al. 2020

Preprint

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Background: Toxoplasma gondii is a single-celled parasite commonly found in mammals. Diagnosis of toxoplasmosis largely depends on measurements of the antibody and/or antigen and Toxoplasma-derived DNAs due to the presence of tissue dwelling quiescent cysts and latent infection of the parasite. As a major surface antigen of T.gondii tachyzoites, SAG1 is a key marker for laboratory diagnosis. However, at present, there are no methods available for SAG1 detection using aptamer-based technology.Methods: Recombinant truncated SAG1（r-tSAG1）of Toxoplasma WH3 strain (type Chinese 1) was prokaryotically expressed and subjected to the synthetic oligonucleotide library for selection of nucleic acid aptamer which targets the r-tSAG1, with systematic evolution of ligands by exponential enrichment (SELEX) strategy. The screened specific aptamer-2 was used in direct enzyme-linked aptamer assay (DELAA) to detect native SAG1 obtained from tachyzoite lysates, mouse sera of acute infection, and human sera that had been verified to be positive for ToxoDNAs by PCR amplification. Results: The soluble r-tSAG1 protein was obtained from E.coli lysates by using 0.01M Tris-Cl in PBS, and was purified and identified by immunoblotting, and then labelled with biotin. The screened aptamers were amplified by PCR, followed by DNA sequencing. The results showed that the aptamer-2, with the highest affinity to nSAG1 among the four aptamer candidates, has a higher specificity and sensitivity when used in detection of nSAG1 in the sera of both animals and humans when compared with the commercial Toxoplasma circulating antigen testing kit.Conclusions: A new direct enzyme-linked aptamer assay (DELAA), with aptamer-2 as the recognition probe, was developed for detection of native SAG1 protein secreted by T.gondii. With increased sensitivity and specificity, stability during storage, easy and cheaper production, the aptamer-based technique is considered as a efficient method for the diagnosis of active and reactivated toxoplasmosis.

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Section: Discussionmentioning

confidence: 99%

SELEX-Based Direct Enzyme-Linked Aptamer Assay(DELAA) for Diagnosis of Toxoplasmosis by Detection of SAG1 Antigen in Sera of Mice and Humans

Shen

Cui

Cudjoe

et al. 2020

Preprint

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“…Another aptamer, MTB36 also shows preferential binding with M.tb over M . bovis BCG suggesting its ability to discriminate between close species ( Mozioglu et al., 2016 ). PPK (polyphosphate kinase gene) regulates inorganic polyphosphate (polyP) function and aids in virulence.…”

Section: Aptamers In the Diagnostic And Therapeutic Applications Of Mtbmentioning

confidence: 99%

Aptamers: An Emerging Tool for Diagnosis and Therapeutics in Tuberculosis

Srivastava

Abraham

Mukhopadhyay

2021

Front. Cell. Infect. Microbiol.

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Tuberculosis (TB) has been plaguing human civilization for centuries, and currently around one-third of the global population is affected with TB. Development of novel intervention tools for early diagnosis and therapeutics against Mycobacterium tuberculosis (M.tb) is the main thrust area in today’s scenario. In this direction global efforts were made to use aptamers, the chemical antibodies as tool for TB diagnostics and therapeutics. This review describes the various aptamers introduced for targeting M.tb and highlights the need for development of novel aptamers to selectively target virulent proteins of M.tb for vaccine and anti-TB drugs. The objective of this review is to highlight the diagnostic and therapeutic application of aptamers used for tuberculosis. The discovery of aptamers, SELEX technology, different types of SELEX development processes, DNA and RNA aptamers reported for diseases and pathogenic agents as well have also been described in detail. But the emphasis of this review is on the development of aptamers which can block the function of virulent mycobacterial components for developing newer TB vaccine candidates and/or drug targets. Aptamers designed to target M.tb cell wall proteins, virulent factors, secretory proteins, or combination could orchestrate advanced diagnosis and therapeutic measures for tuberculosis.

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“…Multiple aptamers have been developed that target strains of this bacterium or virulence factors for its havoc in humans. Mozioglu et al isolated the H37Ra strain as the positive target in their SELEX study [ 33 ]. Both ultrafiltration and centrifugation methods were used to partition the bound and unbound library molecules.…”

Section: In Vitro Selection Of Ssdna Aptamersmentioning

confidence: 99%

Recent Progress in the Identification of Aptamers Against Bacterial Origins and Their Diagnostic Applications

Trunzo

Hong

2020

IJMS

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Aptamers have gained an increasing role as the molecular recognition element (MRE) in diagnostic assay development, since their first conception thirty years ago. The process to screen for nucleic acid-based binding elements (aptamers) was first described in 1990 by the Gold Laboratory. In the last three decades, many aptamers have been identified for a wide array of targets. In particular, the number of reports on investigating single-stranded DNA (ssDNA) aptamer applications in biosensing and diagnostic platforms have increased significantly in recent years. This review article summarizes the recent (2015 to 2020) progress of ssDNA aptamer research on bacteria, proteins, and lipids of bacterial origins that have implications for human infections. The basic process of aptamer selection, the principles of aptamer-based biosensors, and future perspectives will also be discussed.

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Selection of Nucleic Acid Aptamers Specific for Mycobacterium tuberculosis

Cited by 37 publications

References 54 publications

SELEX-Based Direct Enzyme-Linked Aptamer Assay(DELAA) for Diagnosis of Toxoplasmosis by Detection of SAG1 Antigen in Sera of Mice and Humans

SELEX-Based Direct Enzyme-Linked Aptamer Assay(DELAA) for Diagnosis of Toxoplasmosis by Detection of SAG1 Antigen in Sera of Mice and Humans

Aptamers: An Emerging Tool for Diagnosis and Therapeutics in Tuberculosis

Recent Progress in the Identification of Aptamers Against Bacterial Origins and Their Diagnostic Applications

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