Rapid diagnosis of tuberculosis through the detection of mycobacterial DNA in urine by nucleic acid amplification methods

Green, Clare; Huggett, Jim F.; Talbot, Elizabeth A.; Mwaba, Peter; Reither, Klaus; Zumla, Alimuddin

doi:10.1016/s1473-3099(09)70149-5

Cited by 107 publications

(114 citation statements)

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“…During infection, Mycobacterium-specific proteins, peptides, DNA, and lipids are released into the circulation (14,15) and are likely to provide unprecedented specificity to identify M. bovis infection. We hypothesized that a panel of biomarkers, including host and pathogen peptides, would establish a precise test to detect subclinical bTB infection.…”

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confidence: 99%

Circulating Mycobacterium bovis Peptides and Host Response Proteins as Biomarkers for Unambiguous Detection of Subclinical Infection

et al. 2014

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e Bovine tuberculosis remains one of the most damaging diseases to agriculture, and there is also a concern for human spillover. A critical need exists for rapid, thorough, and inexpensive diagnostic methods capable of detecting and differentiating Mycobacterium bovis infection from other pathogenic and environmental mycobacteria at multiple surveillance levels. In a previous study, Seth et al. (PLoS One 4:e5478, 2009, doi:10.1371/journal.pone.0005478) identified 32 host peptides that specifically increased in the blood serum of M. bovis-infected animals). In the current study, 16 M. bovis proteins were discovered in the blood serum proteomics data sets. A large-scale validation analysis was undertaken for selected host and M. bovis proteins using a cattle serum repository containing M. bovis (n ‫؍‬ 128), Mycobacterium kansasii (n ‫؍‬ 10), and Mycobacterium avium subsp. paratuberculosis (n ‫؍‬ 10), cases exposed to M. bovis (n ‫؍‬ 424), and negative controls (n ‫؍‬ 38). Of the host biomarkers, vitamin D binding protein (VDBP) showed the greatest sensitivity and specificity for M. bovis detection. Circulating M. bovis proteins, specifically polyketide synthetase 5, detected M. bovis-infected cattle with little to no seroreactivity against M. kansasii-and M. avium subsp. paratuberculosis-infected animals. These data indicate that host and pathogen serum proteins can serve as reliable biomarkers for tracking M. bovis infection in animal populations.

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confidence: 99%

Circulating Mycobacterium bovis Peptides and Host Response Proteins as Biomarkers for Unambiguous Detection of Subclinical Infection

et al. 2014

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“…19 Consistent with our findings of increased sensitivity with retention of urine supernatant, previous authors have documented that this contains transrenal DNA, which may be beneficial in increasing detection of TB DNA. 6,17 The two main gene targets explored in this review include the M. tuberculosis complex-specific insertion sequence, IS6110, which has been suggested to be the standard tool for TB diagnosis; 32 and the gene coding for the β subunit of the bacterial RNA polymerase, rpoB, employed in Xpert, which amplifies an M. tuberculosis-specific sequence of this gene and detects mutations to determine rifampicin resistance. 33 In the face of an undefined optimal gene target for urine TB NAAT, 9,10,14,15,17,18 our findings suggest that rpoB may be less sensitive than IS6110.…”

Section: Discussionmentioning

confidence: 99%

“…[2][3][4] The relative simplicity of urine collection could be ideal to address diagnostic challenges of active PTB, 5 and could be exploited in point-of-care (POC) testing of active PTB at all levels of health care, and potentially in households and the community. 6,7 In addition to being safer to handle, normal urine production yields frequent and relatively large amounts, making urine specimens convenient to obtain and easier to test in comparison to sputum. Children could benefit the most from urine NAATs for PTB diagnosis due to challenges in obtaining sputum samples, particularly from young children who cannot expectorate, and the paucibacillary nature of their disease.…”

Section: Introductionmentioning

confidence: 99%

“…9,10 Cell-free nucleic acids released from dying human cells and Mycobacterium tuberculosis microorganisms may be filtered through the kidneys, resulting in the detection of trans-renal DNA fragments in urine. 6 Although the exact pathophysiological mechanisms underlying the detection of mycobacterial DNA in the urine of patients with active PTB are not well understood, it has high potential relevance for clinical practice.The objective of this systematic review and meta-analysis was to determine the diagnostic accuracy of M. tuberculosis NAATs in urine samples for active PTB compared to sputum culture as the reference standard. …”

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Diagnostic accuracy of nucleic acid amplification tests in urine for pulmonary tuberculosis: A meta-analysis

2015

Indian Journal of Tuberculosis

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OBJECTIVE-To determine the diagnostic accuracy of tuberculosis (TB) nucleic acid amplification tests (NAATs) in urine samples for individuals with active pulmonary tuberculosis (PTB).DESIGN-Systematic review and meta-analysis. Electronic databases and reference lists were searched without age or setting restrictions up to May 2015. Eligible articles examined Mycobacterium tuberculosis NAATs in urine samples for PTB diagnosis in patients with sputum culture as the reference standard, and reported sufficient data to separately calculate sensitivity or specificity.RESULTS-Eight studies, including 1212 participants from seven countries with a mean age ranging from 28 to 48 years, were included. Polymerase chain reaction (PCR) with insertion sequence (IS) 6110, rpoB or cfp32/hf6 as gene targets was used for NAATs. The pooled sensitivity and specificity was respectively 0.55 (95%CI 0.36-0.72) and 0.94 (95%CI 0.78-0.99), with slightly higher sensitivity in human immunodeficiency virus positive individuals, at 0.59 (95%CI 0.20-0.89). Sensitivity was higher in sputum microscopy-positive than -negative individuals. Storage temperatures below −70°C, centrifuge speed <5000 rpm and IS6110 increased sensitivity on meta-regression.Correspondence to: Diana Marangu, Department of Paediatrics and Child Health, University of Nairobi, P O Box 19676 -0202, Nairobi, Kenya. marangud@gmail.com. Conflicts of interest: none declared. HHS Public Access KeywordsNAAT; PCR; systematic review; meta-regression In 2013, it was estimated that there were 9 million incident tuberculosis (TB) cases globally, 3.3 million of whom were not diagnosed. 1 Urine nucleic acid amplification tests (NAATs) have potential for use in the diagnosis of pulmonary tuberculosis (PTB), the most common form of TB worldwide. [2][3][4] The relative simplicity of urine collection could be ideal to address diagnostic challenges of active PTB, 5 and could be exploited in point-of-care (POC) testing of active PTB at all levels of health care, and potentially in households and the community. 6,7 In addition to being safer to handle, normal urine production yields frequent and relatively large amounts, making urine specimens convenient to obtain and easier to test in comparison to sputum. Children could benefit the most from urine NAATs for PTB diagnosis due to challenges in obtaining sputum samples, particularly from young children who cannot expectorate, and the paucibacillary nature of their disease. 5,8 With the advancement of molecular techniques and wider use of the NAAT Xpert ® MTB/RIF assay (Cepheid, Sunnyvale, CA, USA) in many high TB endemic countries, exploration of alternative samples such as urine for TB diagnosis is increasing due to its ease of collection and potential for decentralized implementation. 9,10 Cell-free nucleic acids released from dying human cells and Mycobacterium tuberculosis microorganisms may be filtered through the kidneys, resulting in the detection of trans-renal DNA fragments in urine. 6 Although the exact pathophysiological mech...

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“…Most of these discrepancies have been attributed to differences in the methods used for collection and storage of urine specimens and in the extraction and amplification of Tr-DNA. It has been suggested that improvement of extraction techniques, targeting Tr-DNA fragments smaller than 150 to 200 bp, and the use of real-time PCR might increase sensitivity and specificity of the test (8). In our study, we employed a magnetic-bead-based extraction method which is effective for extracting 200-bp fragments and therefore suitable for detecting Tr-DNA (9).…”

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Use of Transrenal DNA for the Diagnosis of Extrapulmonary Tuberculosis in Children: a Case of Tubercular Otitis Media

et al. 2015

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The diagnosis of tuberculosis (TB) is difficult in children, especially for smear-negative pulmonary and extrapulmonary TB, which are common at this age. We report an 11-year-old girl with TB otitis media with negative smear microscopy and Xpert MTB/RIF but positive Mycobacterium tuberculosis-specific transrenal DNA (Tr-MTB-DNA) test results and culture for M. tuberculosis. CASE REPORTA n 11-year-old girl was admitted to the Department of Otolaryngology of the S. Orsola-Malpighi University Hospital, Bologna, Italy, with a 1-year history of bilateral chronic otitis media associated with conductive hearing loss; she had received several courses of broad-spectrum antibiotic therapy without significant improvement. A myringotomy was planned to aspirate the exudate and decompress the tympanic cavity, but instead of the expected effusion, a diffuse tympanic granulomatosis was noted. The child was therefore referred to the Pediatric Respiratory Service for further investigation to exclude a diagnosis of tubercular otitis media (TBOM). The child had a medical history of asthma and dust allergy but was otherwise healthy. Her parents had originated in Bangladesh, but she was born and had grown up in Italy. She had traveled to Bangladesh for 6 months the year before the beginning of the symptoms but had no known contact with persons having tuberculosis (TB) or chronic cough.On admission, the child was in good condition, her general examination was normal, and there was no evidence of other disease localizations or disseminated TB. A chest X-ray was unremarkable. A tuberculin skin test (TST) (25-mm induration) and Quantiferon-TB Gold in-tube (QFT-IT; Qiagen, Germany; TBAg-Nil ϭ 1.46 IU interferon gamma/ml) test were positive. In addition to the auricular secretion collected during micro-otoscopy, a multiple-sampling approach was performed: nasopharyngeal aspirates (NPA), gastric aspirates (GA), and sputum and urine samples were collected on consecutive days. Each sample underwent direct smear microscopy after Ziehl-Neelsen staining and testing with an Xpert MTB/RIF assay (Cepheid, France), and all were negative. All the specimens were also cultured on liquid (Bactec MGIT 960; Becton Dickinson, USA) and solid (Lowenstein-Jensen; Heipha, Germany) media.In order to detect Mycobacterium tuberculosis-specific transrenal DNA (Tr-MTB-DNA), as described by Cannas et al.(1), 6 urine samples (50 ml each) were collected during the study period, as depicted in Fig. 1, and quickly added to 1 ml of a stabilization buffer (0.5 M EDTA-0.5 M Tris-HCl, pH 8.5) to a final concentration of 10 mM, in order to minimize degradation of soluble DNA by nucleases. The stabilized urine specimens were then stored in 1-ml aliquots at Ϫ80°C. DNA was extracted from 1 ml of whole urine and eluted in 25 l with an automated system based on magnetic silica particle technology (NucliSENS easyMAG; bioMérieux, France); urine specimens had previously been lysed offboard at 95°C for 30 min. Tr-MTB-DNA was amplified by inhouse seminested PCR, targeting the M. tuber...

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Rapid diagnosis of tuberculosis through the detection of mycobacterial DNA in urine by nucleic acid amplification methods

Cited by 107 publications

References 55 publications

Circulating Mycobacterium bovis Peptides and Host Response Proteins as Biomarkers for Unambiguous Detection of Subclinical Infection

Circulating Mycobacterium bovis Peptides and Host Response Proteins as Biomarkers for Unambiguous Detection of Subclinical Infection

Diagnostic accuracy of nucleic acid amplification tests in urine for pulmonary tuberculosis: A meta-analysis

Use of Transrenal DNA for the Diagnosis of Extrapulmonary Tuberculosis in Children: a Case of Tubercular Otitis Media

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