Application of Droplet Digital PCR to Detection of Mycobacterium tuberculosis and Mycobacterium leprae Infections: A Narrative Review

Fan, Yuxin; Chen, Jingjing; Liu, Meixiao; Xu, Xin; Yue, Peng; Cao, Wenjing; Ji, Zhenhua; Su, Xuan; Wen, Shiyuan; Kong, Jing; Zhou, Guozhong; Li, Bingxue; Dong, Yan; Liu, Aihua; Bao, Fukai

doi:10.2147/idr.s349607

Cited by 7 publications

(12 citation statements)

References 59 publications

(87 reference statements)

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“…The sensitivity and specificity of the mpt64 assay using RT-PCR to detect MTB from the peripheral blood were 30.8% and 88.8%, respectively [ 35 ]. ddPCR is a highly sensitive and specific method for nucleic acid quantification that has no need for a calibration curve [ 14 ]. The sensitivity and specificity of the mpt64 antigen test were 92% and 95%, respectively [ 36 ].…”

Section: Discussionmentioning

confidence: 99%

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Detection of Mycobacterium tuberculosis Complex in Sputum Samples Using Droplet Digital PCR Targeting mpt64

et al. 2023

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Tuberculosis (TB) is one of the top 10 causes of death worldwide. It is challenging to find methods of diagnosis of active pulmonary TB that are sensitive enough to detect cases for proper treatment before unintentional transmission. Droplet digital PCR (ddPCR) is a highly sensitive method to detect genetic material of pathogens, but it has rarely been used for diagnosis of TB. This study compared the sensitivity of ddPCR with that of GeneXpert and AFB smear microscopy in 180 leftover sputum samples from patients suspected of having TB on the basis of clinical symptoms and radiography. Absolute quantification of copy numbers of MTB-specific genes was possible using ddPCR targeting the mpt64 gene. Among the 180 samples, 41.1% were diagnosed as having TB using ddPCR. The sensitivities of AFB smear microscopy, GeneXpert and ddPCR were 41.9%, 82.4% and 100%, respectively. AFB smear microscopy and GeneXpert both had a specificity of 100%, and the specificity of ddPCR was 95.3%. The accuracy of ddPCR (97.2%) is higher than that of GeneXpert (92.7%). This robust ddPCR system could potentially be used as a method for early diagnosis of TB.

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Section: Discussionmentioning

confidence: 99%

“…Additional advantages are absolute quantification, unparalleled precision, superior partitioning and the best accuracy in the presence of PCR inhibitors [ 13 ]. This technology has rarely been used for TB diagnosis [ 14 ] and its performance has never been investigated in a country with a high TB burden.…”

Section: Introductionmentioning

confidence: 99%

Detection of Mycobacterium tuberculosis Complex in Sputum Samples Using Droplet Digital PCR Targeting mpt64

et al. 2023

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“…Digital droplet PCR (ddPCR) is one of the emerging PCR, reported in various fields of research, including food adulterants and drug testing; detection of viral, parasite, or bacterial pathogens; gene modification; and clinical diagnosis (Fan et al., 2022; Kim et al., 2020; Luo et al., 2022; Ren et al., 2017). Compared to qPCR, its main advantage is the ability to quantify nucleic acids without the need for a standard curve.…”

Section: Nucleic Acid‐based Amplification Methods For Targeted Detect...mentioning

confidence: 99%

Molecular detection and characterization of foodborne bacteria: Recent progresses and remaining challenges

Hadi

Rapp

Dhawan

et al. 2023

Comp Rev Food Sci Food Safe

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The global food demand is expected to increase in the coming years, along with challenges around climate change and food security. Concomitantly, food safety risks, particularly those related to bacterial pathogens, may also increase. Thus, the food sector needs to innovate to rise to this challenge. Here, we discuss recent advancements in molecular techniques that can be deployed within various foodborne bacteria surveillance systems across food settings. To start with, we provide updates on nucleic acid-based detection, with a focus on polymerase chain reaction (PCR)-based technologies and loop-mediated isothermal amplification (LAMP). These include descriptions of novel genetic markers for several foodborne bacteria and progresses in multiplex PCR and droplet digital PCR.The next section provides an overview of the development of clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated (Cas) proteins systems, such as CRISPR-Cas9, CRISPR-Cas12a, and CRISPR-Cas13a, as tools for enhanced sensitive and specific detection of foodborne pathogens. The final section describes utilizations of whole genome sequencing for accurate characterization of foodborne bacteria, ranging from epidemiological surveillance to model-based predictions of bacterial phenotypic traits through genome-wide association studies or machine learning.

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“…Detection of Infections ddPCR [91,92] Identification of risk genes that affect susceptibility to leprosy NSG [93]…”

Section: Mycobacterium Lepraementioning

confidence: 99%

Metabarcoding and Digital PCR (dPCR): Application in the Study of Neglected Tropical Diseases

Lagunes-Castro¹,

López‐Monteon²,

Guzmán-Gómez³

et al. 2023

New Advances in Neglected Tropical Diseases

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neglected tropical diseases such as Chagas disease, dengue, Zika, chikungunya, and malaria cause millions of deaths each year and they are caused by a variety of pathogens whose diagnosis is very limited or subject to conventional testing, making a treatment less accessible, accurate and timely diagnosis for choosing their treatments. Traditional methods for pathogen detection have not been able to meet the growing need for diagnosis and control. The incorporation of new technologies such as next-generation sequencing (NGS) and digital PCR (dPCR) represent a better diagnostic possibility due to their ability to absolutely quantify pathogens with high selectivity and precision. Our planet is currently experiencing environmental changes of an unprecedented magnitude and rate, including climate change, globalized pollution, biodiversity loss, and land use changes, so neglected diseases require a comprehensive understanding of the ecology of vectors in the different eco-epidemiological contexts, as well as of the transmission cycles of pathogens and their transmission dynamics. In this sense, NGS and dPCR open a new panorama for a better understanding of these diseases with the aim of proposing new programs for their care.

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Application of Droplet Digital PCR to Detection of Mycobacterium tuberculosis and Mycobacterium leprae Infections: A Narrative Review

Cited by 7 publications

References 59 publications

Detection of Mycobacterium tuberculosis Complex in Sputum Samples Using Droplet Digital PCR Targeting mpt64

Detection of Mycobacterium tuberculosis Complex in Sputum Samples Using Droplet Digital PCR Targeting mpt64

Molecular detection and characterization of foodborne bacteria: Recent progresses and remaining challenges

Metabarcoding and Digital PCR (dPCR): Application in the Study of Neglected Tropical Diseases

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