A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis

Zhao, Zhenzhen; Wu, T.; Wang, Minjin; Chen, Xiaojuan; Liu, Tangyuheng; Si, Yanjun; Zhou, Yanhong; Ying, Binwu

doi:10.1016/j.ijid.2022.07.041

Cited by 7 publications

(4 citation statements)

References 29 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…More smear-positive samples are needed to fully evaluate the kit’s performance in identifying the MDR. On the other hand, a modified method such as droplet digital PCR and multiplex allele-specific PCR could also improve the sensibility of mutation gene detection ( 38 – 40 ).…”

Section: Discussionmentioning

confidence: 99%

Evaluation of mycobacteria infection prevalence and optimization of the identification process in North Sardinia, Italy

Chen,

Sechi,

Molicotti

2024

Microbiol Spectr

View full text Add to dashboard Cite

Mycobacterium tuberculosis (MTB) and non-tuberculous mycobacteria (NTM) are the most common pathogens of mycobacterial infection, and their prevalence varies around the world. This study aimed to estimate the prevalence of mycobacterial infection and drug resistance in North Sardinia, Italy and to optimize the process of mycobacteria identification. From January 2020 to April 2023, samples from 1,836 suspected individuals in North Sardinia were analyzed. Conventional methods (microscopy smear and culture) and molecular methods (polymerase chain reaction, DNA probe, and mass spectrum) were used in MTB and NTM identification. Among 1,836 suspected individuals, 89 were MTB-positive and 44 were NTM-positive. Bronchial aspiration and bronchoalveolar lavage performed well as substitute specimens for sputum in MTB detection. For drug resistance, 71.91% of MTB and 83.33% of NTM were resistant to at least one antibiotic. The average turn-around time of MTB, slowly growing mycobacteria, and rapidly growing mycobacteria was 7.12, 48.87, and 11.27 days, respectively. Overall, our results show that the prevalence of mycobacterial infection in North Sardinia is relatively optimistic. However, the high proportion of drug resistance detected in the isolates is alarming. The implementation of advanced techniques could help optimize the process of mycobacteria identification. IMPORTANCE Mycobacterial infection is a major threat to public health worldwide. Accurate identification of infected species and drug resistance detection are critical factors in treatment. We focused on shortening the turn-around time of identifying mycobacteria species and antibiotic resistance tests.

show abstract

Section: Discussionmentioning

confidence: 99%

Evaluation of mycobacteria infection prevalence and optimization of the identification process in North Sardinia, Italy

Chen,

Sechi,

Molicotti

2024

Microbiol Spectr

View full text Add to dashboard Cite

show abstract

“…ddPCR, which is a method that quantifies the absolute copy number of a target DNA, is more sensitive and repeatable than real-time PCR ( 18 , 19 ) for the detection of the pathogen K. pneumoniae in fecal samples; ddPCR may provide a more simple and sensitive technique. ddPCR has been widely applied to the quantification of various pathogens, such as Mycobacterium turberculosis ( 20 , 21 ), Ureaplasma spp. ( 22 ), Verticillium dahliae , Verticillium longisporum ( 23 ), hepatitis B virus ( 24 ), and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ( 25 ).…”

Section: Discussionmentioning

confidence: 99%

Detection and Quantification of Klebsiella pneumoniae in Fecal Samples Using Digital Droplet PCR in Comparison with Real-Time PCR

Feng

Cui

et al. 2023

Microbiol Spectr

View full text Add to dashboard Cite

show abstract

“…Some suspected tuberculosis diagnoses still lacked etiological evidence. ddPCR is an excellent nucleic acid quantification technique with high sensitivity (Zhao et al, 2022). However, directly evaluating the diagnostic performance of ddPCR with mNGS and Xpert in pulmonary MTB infection is still rare so far.…”

Section: Discussionmentioning

confidence: 99%

ddPCR provides a sensitive test compared with GeneXpert MTB/RIF and mNGS for suspected Mycobacterium tuberculosis infection

Zhang,

Yu,

Han

et al. 2023

Front. Cell. Infect. Microbiol.

View full text Add to dashboard Cite

IntroductionThe Metagenomics next-generation sequencing (mNGS) and GeneXpert MTB/RIF assay (Xpert) exhibited a sensitivity for tuberculosis (TB) diagnostic performance. Research that directly compared the clinical performance of ddPCR analysis, mNGS, and Xpert in mycobacterium tuberculosis complex (MTB) infection has not been conducted.MethodsThe study aimed to evaluate the diagnostic performance of ddPCR compared to mNGS and Xpert for the detection of MTB in multiple types of clinical samples. The final clinical diagnosis was used as the reference standard.ResultsOut of 236 patients with suspected active TB infection, 217 underwent synchronous testing for tuberculosis using ddPCR, Xpert, and mNGS on direct clinical samples. During follow-up, 100 out of 217 participants were diagnosed with MTB infection. Compared to the clinical final diagnosis, ddPCR produced the highest sensitivity of 99% compared with mNGS (86%) and Xpert (64%) for all active MTB cases. DiscussionTwenty-two Xpert-negative samples were positive in mNGS tests, which confirmed the clinical diagnosis results from ddPCR and clinical manifestation, radiologic findings. Thirteen mNGS-negative samples were positive in ddPCR assays, which confirmed the clinical final diagnosis.ddPCR provides a higher sensitive compared to Xpert and mNGS for MTB diagnosis, as defined by the high concordance between ddPCR assay and clinical final diagnosis.

show abstract

A new droplet digital PCR assay: improving detection of paucibacillary smear-negative pulmonary tuberculosis

Cited by 7 publications

References 29 publications

Evaluation of mycobacteria infection prevalence and optimization of the identification process in North Sardinia, Italy

Evaluation of mycobacteria infection prevalence and optimization of the identification process in North Sardinia, Italy

Detection and Quantification of Klebsiella pneumoniae in Fecal Samples Using Digital Droplet PCR in Comparison with Real-Time PCR

ddPCR provides a sensitive test compared with GeneXpert MTB/RIF and mNGS for suspected Mycobacterium tuberculosis infection

Contact Info

Product

Resources

About