Evaluation of a simple loop-mediated isothermal amplification test kit for the diagnosis of tuberculosis

Mitarai, Satoshi; Okumura, Meinoshin; Toyota, Emiko; Yoshiyama, Takashi; Ando, Akio; Sejimo, Akiko; Azuma, Yuka; Sugahara, Kazunori; Nagasawa, Toshiro; Nagayama, Naohiro; Yamane, Akira; Yano, Ryozo; Kokuto, Hiroyuki; Morimoto, Kozo; Ueyama, Masashi; Kubota, Motoko; Ren, Yi; Ogata, Hideo; Kudoh, S; Mori, Toru

doi:10.5588/ijtld.10.0629

Cited by 108 publications

(100 citation statements)

References 28 publications

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“…За препарат-позитивне узорке спутума осјетљивост је била 73.6%, а за препарат-негативне 52.1% (Fegou et al, 2005). Осјетљивост COBAS AMPLICOR MTB Теста код препарат-позитивних респираторних узорака у истраживању које су провели Mitarai et al износила је 99.4%, потврђујући оправданост примјене теста за брзу дијагнозу ТB код ових узорака (Mitarai et al, 2011). Осјетљивост примјењеног COBAS AMPLICOR MTB Теста у овом раду је била 97.87% код препарат-позитивних узорака спутума, док је код препарат-негативних узорака спутума износила 80%.…”

Section: дискусијаunclassified

РЕТРОСПЕКТИВНА ЕВАЛУАЦИЈА COBAS AMPLICOR MTB ТЕСТА У ДЕТЕКЦИЈИ Mycobacterium tuberculosis ИЗ РЕСПИРАТОРНИХ УЗОРАКА

Халиловић¹,

Нумановић²,

Тихић³

et al. 2016

SBERS

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Traditional methods for detection of mycobacteria are limited by low sensitivity and/or specificity, with the time necessary for the identification of Mycobacterium tuberculosis of several weeks. A number of molecular techniques, including different biomarkers, have been developed for rapid identification of mycobacteria. The aim of this study was to evaluate the reliability of the COBAS AMPLICOR MTB Test in detection of Mycobacterium tuberculosis in respiratory specimens. Samples were decontaminated with N-acetyl-L-cysteine-sodium hydroxide (NALC-NaOH). One drop was used for detection of acid-fast bacilli positive-smears by Ziehl-Neelsen (ZN) staining, 200 µL to inoculate Löwenstein-Jensen (LJ) medium, 500 µL for cultivation in BACTEC MGIT 960 System, and 100 µL for the COBAS AMPLICOR MTB Test, performed according to manufacturer's recommendations. Out of 100 samples, 50 samples were smear-positive and 50 smear-negative. Mycobacterium tuberculosis was isolated from 59 samples by at least one cultivation method. After the analysis of inconsistent results, the overall sensitivity, specificity, positive and negative predictive value of the COBAS AMPLICOR MTB Test were 94.93%, 100%, 100% and 91.89% compared to diagnostic cultures, with an inhibition rate 2%. The values for smear-positive and smear-negative samples were 97.87%, 100%, 100%, 50%, and 83.33%, 100%, 100% and 94.29%, respectively. The greater numbers of positive samples were detected using the COBAS AMPLICOR MTB Test than by direct microscopy. Based on the obtained results, considering the methodological simplicity and completely automatized amplification and detection, the COBAS AMPLICOR MTB Test has proven to be very reliable in the diagnosis of tuberculosis (TB).

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Section: дискусијаunclassified

РЕТРОСПЕКТИВНА ЕВАЛУАЦИЈА COBAS AMPLICOR MTB ТЕСТА У ДЕТЕКЦИЈИ Mycobacterium tuberculosis ИЗ РЕСПИРАТОРНИХ УЗОРАКА

Халиловић¹,

Нумановић²,

Тихић³

et al. 2016

SBERS

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“…For example, a commercially available ultrarapid DNA extraction method (PURE; Eiken) could be used to improve the sensitivity. In this case, the homogenization steps followed by centrifugation can be omitted and this would prevent the loss of bacteria during the process, as reported by Mitarai et al (2011).From a public health and infection control viewpoint, the LAMP method has an advantage in the clinical laboratory as it permits immediate distinction of MTC infection (with the need for patient quarantine) from NTM infection (without such need for quarantine). Conventionally, the diagnosis of TB at the hospital is based on smear microscopy, X-ray and culture confirmation.…”

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confidence: 93%

“…Although several clinical evaluations of LAMP assays have been published (Boehme et al, 2007;George et al, 2011;Mitarai et al, 2011;Pandey et al, 2008), to the best of our knowledge, this is the first study to clinically evaluate the use of the LAMP method to detect the hspX gene of MTC and is also the first report of the LAMP method for diagnosing TB in Korea. Sputum specimens obtained from patients at the Korea University Medical Center (KUMC) who were suspected of having TB were subjected to culture confirmation, smear microscopy and the LAMP method.…”

Section: Introductionmentioning

confidence: 99%

“…From 3 September to 21 December 2012, a total of 303 sputum specimens were obtained from patients with suspected TB at the KUMC. Using the obtained specimens, an experienced laboratory technician performed smear microscopy according to the acid-fast bacillus stains, including auramine-O and Ziehl-Neelsen stainings, as described previously (Hänscheid et al, 2007).The clinical specimens were also processed as described previously (Mitarai et al, 2011). Briefly, an equal volume of decontaminationdigestion agent, NALC (N-acetyl-L-cysteine)/2 % NaOH, was added to the sputum in a 50 ml falcon tube (BD Diagnostics).…”

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confidence: 99%

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Detection of Mycobacterium tuberculosis complex in sputum specimens using a loop-mediated isothermal amplification assay in Korea

Moon

Kim

Tomono

et al. 2015

Journal of Medical Microbiology

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Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis complex (MTC), remains one of the leading causes of death in the world. In Korea, the current prevalence of multidrug-resistant TB (MDR-TB) poses a major problem. The most common method for diagnosing TB in developing countries is sputum smear microscopy; however, the sensitivity of this test is relatively low and it usually requires well-trained laboratory staff. Cultures of MTC require up to several weeks in sophisticated facilities, such as Biosafety Level 3. Effective diagnostic techniques are necessary to control TB. In Korea, we evaluated a loop-mediated isothermal amplification (LAMP) assay targeting the hspX gene (TB-hspX-LAMP) of MTC. For clinical evaluation, culture confirmation, smear microscopy and TB-hspX-LAMP were performed on 303 sputum specimens obtained from suspected TB patients in Korea. The sensitivity, specificity, positive predictive value and negative predictive value of TB-hspX-LAMP were 71.1, 98.8, 91.4 and 95.1 %, respectively, compared with TB culture, which is the gold standard for diagnosis of TB. In contrast, the comparable values of smear microscopy were 24.4, 98.1, 68.8 and 88.2 %, respectively. Therefore, we concluded that TB-hspX-LAMP was superior to the use of smear microscopy for the detection of MTC in sputum specimens in clinical settings in Korea. INTRODUCTIONTuberculosis (TB), an airborne infectious disease caused by Mycobacterium tuberculosis complex (MTC, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium africanum, Mycobacterium microti, Mycobacterium caprae, Mycobacterium canetti, Mycobacterium pinnipedii) (Behr et al., 1999; CDC, 2009), still remains one of the leading causes of death by infectious diseases worldwide. In 2013, 9 million people developed TB throughout the world; 1.5 million of this group died from it, of which 360 000 were human immunodeficiency virus-positive (WHO, 2014).Early, accurate diagnosis of MTC is important for the control of TB (Brodie & Schluger, 2005). The most common method for diagnosing TB in developing countries is sputum smear microscopy (George et al., 2011). Smear microscopy offers the advantages of rapid detection time and simplicity. However, the sensitivity of smear microscopy is relatively low and it requires well-trained laboratory staff.Culture methods can detect as few as 100 Mycobacterium cells per 1 ml specimen (Hofmann-Thiel et al., 2010) and TB culture is still considered the gold standard for the et al., 2003). The LAMP reaction produces stem-loop DNA structures with several inverted repeats and cauliflower-like structures with multiple loops. The products of amplified LAMP reactions can be measured by a change in the turbidity of the reaction mixture because magnesium pyrophosphate accumulates in the reaction mixture as a byproduct of the amplification reaction.The LAMP assay has several exceptional features compared with other TB diagnostic tools.(1) The LAMP assay uses four different primers to identify six distinct regions on...

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“…Three main technical aspects of the test are (a) amplification of DNA with LAMP, (b) ultra-rapid extraction (PURE) of DNA that can remove inhibitors of the test without significant loss of genetic material and (c) straightforward visual detection of the result using fluorescence under ultra-violet light. 98,99 The Loopamp TM LF-160 machine does not need maintenance or calibration which makes the overall process simple and easy to pick up; dismissing the need for extensively trained personnel. The sensitivity of TB LAMP is 100 times higher than sputum smear microscopy.…”

Section: Host Responses To M Tuberculosismentioning

confidence: 99%

Early diagnosis and effective treatment regimens are the keys to tackle antimicrobial resistance in tuberculosis (TB): A report from Euroscicon's international TB Summit 2016

et al. 2016

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To say that tuberculosis (TB) has regained a strong foothold in the global human health and wellbeing scenario would be an understatement. Ranking alongside HIV/AIDS as the top reason for mortality due to a single infectious disease, the impact of TB extends far into socio-economic context worldwide. As global efforts led by experts and political bodies converge to mitigate the predicted outcome of growing antimicrobial resistance, the academic community of students, practitioners and researchers have mobilised to develop integrated, inter-disciplinary programmes to bring the plans of the former to fruition. Enabling this crucial requirement for unimpeded dissemination of scientific discovery was the TB Summit 2016, held in London, United Kingdom. This report critically discusses the recent breakthroughs made in diagnostics and treatment while bringing to light the major hurdles in the control of the disease as discussed in the course of the 3-day international event. Conferences and symposia such as these are the breeding grounds for successful local and global collaborations and therefore must be supported to expand the understanding and outreach of basic science research.

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Evaluation of a simple loop-mediated isothermal amplification test kit for the diagnosis of tuberculosis

Abstract: This test is considered a promising diagnostic tool for TB, even for peripheral laboratories with limited equipment, such as those in developing countries.

Cited by 108 publications

References 28 publications

РЕТРОСПЕКТИВНА ЕВАЛУАЦИЈА COBAS AMPLICOR MTB ТЕСТА У ДЕТЕКЦИЈИ Mycobacterium tuberculosis ИЗ РЕСПИРАТОРНИХ УЗОРАКА

РЕТРОСПЕКТИВНА ЕВАЛУАЦИЈА COBAS AMPLICOR MTB ТЕСТА У ДЕТЕКЦИЈИ Mycobacterium tuberculosis ИЗ РЕСПИРАТОРНИХ УЗОРАКА

Detection of Mycobacterium tuberculosis complex in sputum specimens using a loop-mediated isothermal amplification assay in Korea

Early diagnosis and effective treatment regimens are the keys to tackle antimicrobial resistance in tuberculosis (TB): A report from Euroscicon's international TB Summit 2016

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