Improvement of Mycobacterium tuberculosis detection in sputum using DNA extracted by sonication

Bello, Graziele Lima; Morais, Franciele Costa Leite; Wolf, Jonas Michel; Gehlen, Mirela; Soares, Tainá dos Santos; Halon, Maria Laura; Barcellos, Regina Bones; Rossetti, Maria Lúcia Rosa

doi:10.1016/j.bjid.2020.08.006

Cited by 7 publications

(2 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…DNA extraction - The extraction of DNA directly from culture samples and lung samples was performed by the method known as “sonication”. 14 The concentration of DNA extracted from the cultures was determined by spectrophotometry using the Eppendorf BioSpectrometer ® basic equipment (Hamburg, Germany).…”

Section: Methodsmentioning

confidence: 99%

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA

Morais

Bello

Costi

et al. 2022

Mem. Inst. Oswaldo Cruz

Self Cite

View full text Add to dashboard Cite

BACKGROUND Non-tuberculous mycobacteria (NTMs) cause diseases known as mycobacteriosis and are an important cause of morbidity and mortality. The diagnosis of pulmonary disease caused by NTM is hampered by its clinical similarity with tuberculosis (TB) and by the lack of an accurate and rapid laboratory diagnosis. OBJECTIVES Detect DNA from NTMs directly from lung samples using real-time polymerase chain reaction (qPCR) for amplification of 16S rRNA. Additionally, DNA sequencing ( hsp65 and rpoB genes) was used to identify the species of MNTs. METHODS A total of 68 sputum samples (54 with suspected NTMs and 14 with TB) from patients treated at a referral hospital were used. FINDINGS Of these, 27/54 (50%) were qPCR positive for NTMs and 14/14 TB patients (controls) were qPCR negative with an almost perfect concordance ( Kappa of 0.93) with the Mycobacterium spp. culture. Sequencing confirmed the presence of NTM in all positive samples. The most common species was Mycobacterium gordonae (33%), followed by Mycobacterium abscessus (26%), Mycobacterium fortuitum (22%), Mycobacterium avium (15%) and Mycobacterium peregrinum (4%). MAIN CONCLUSIONS The qPCR technique for detecting NTMs targeting 16S rRNA has the potential to detect NTMs and rapidly differentiate from Mycobacterium tuberculosis . However, it is necessary to identify the species to help in the differential diagnosis between disease and contamination, and to guide the choice of the therapeutic scheme.

show abstract

Section: Methodsmentioning

confidence: 99%

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA

Morais

Bello

Costi

et al. 2022

Mem. Inst. Oswaldo Cruz

Self Cite

View full text Add to dashboard Cite

show abstract

“…In the intestinal microflora and compost microbes of fish, the ultrasonic lysis method combined with lysozyme was used to extract DNA ( Yang et al, 2006 ; Han et al, 2018 ). Moreover, direct extraction of DNA from sputum clinical sample (containing diverse microbes) for real-time PCR proved to be convenient and suitable ( Bello et al, 2020 ). In this study, a sonication-based DNA extraction method was developed, which could screen large samples in just one day.…”

Section: Introductionmentioning

confidence: 99%

A Quick DNA Extraction Method for High Throughput Screening in Gram-positive Bacteria

Yuan¹

2023

BIO-PROTOCOL

View full text Add to dashboard Cite

In this study, a sonication-based DNA extraction method was developed, in which the whole process can be finished within 10 min. This method is almost zero cost and time-saving, which is useful for high throughput screening, especially in the screening of mutants generated in random mutagenesis. This method is effective in genomic DNA extraction for PCR amplification in several Gram-positive bacteria, including Bacillus cereus, Bacillus thuringiensis, Bacillus subtilis , and Listeria monocytogenes .

show abstract

Prosthetic joint infection caused by an imipenem-resistant Mycobacterium senegalense

et al. 2023

View full text Add to dashboard Cite

Improvement of Mycobacterium tuberculosis detection in sputum using DNA extracted by sonication

Cited by 7 publications

References 34 publications

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA

Detection of non-tuberculosus mycobacteria (NTMs) in lung samples using 16S rRNA

A Quick DNA Extraction Method for High Throughput Screening in Gram-positive Bacteria

Prosthetic joint infection caused by an imipenem-resistant Mycobacterium senegalense

Contact Info

Product

Resources

About