Diagnostic accuracy of loop-mediated isothermal amplification (LAMP) for detection of Leishmania DNA in buffy coat from visceral leishmaniasis patients

Khan, Gulam Musawwir; Bhaskar, Khondaker Rifat Hasan; Salam, Abdus; Akther, Tania; Pluschke, Gerd; Mondal, Dinesh

doi:10.1186/1756-3305-5-280

Cited by 72 publications

(54 citation statements)

References 29 publications

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“…In contrast, LAMP also demonstrated high sensitivity and specificity in several protozoan studies. Successful amplification of Toxoplasma gondii, Leishmania and Plasmodium falciparum showed a sensitivity of 87.5, 90.7 and 95.8 %, respectively, and a specificity of 100 for all (Lau et al, 2010;Ghayour Najafabadi et al, 2014;Khan et al, 2012). The LAMP assay has also been successfully developed to detect other parasites in different types of clinical samples, such as human saliva, buffy coats, urine and blood (Ghayour Najafabadi et al, 2014;Lau et al, 2011;Khan et al, 2012).…”

Section: Discussionmentioning

confidence: 99%

“…The combination of isothermal conditions and the Bst polymerase in the LAMP assay showed resistance to inhibitors that usually hinder the later stages of conventional PCR (Khan et al, 2012;Mori et al, 2001). LAMP reagents and Bst polymerase are also relatively stable even if kept at 25 and 37 uC (Thekisoe et al, 2009), and only a conventional heating block is required to perform this assay.…”

Section: Discussionmentioning

confidence: 99%

“…LAMP reagents and Bst polymerase are also relatively stable even if kept at 25 and 37 uC (Thekisoe et al, 2009), and only a conventional heating block is required to perform this assay. The requirement for an electrical supply for the heating block can be changed to an alternative power source such as a battery to suit its application (Khan et al, 2012). A water bath could also be used; however, it is recommended that the tubes be wrapped in parafilm to avoid contamination (Khan et al, 2012) as LAMP assays can sometimes lead to false-positive amplification due to the very high specificity and sensitivity (Zhou et al, 2014).…”

Section: Discussionmentioning

confidence: 99%

“…The requirement for an electrical supply for the heating block can be changed to an alternative power source such as a battery to suit its application (Khan et al, 2012). A water bath could also be used; however, it is recommended that the tubes be wrapped in parafilm to avoid contamination (Khan et al, 2012) as LAMP assays can sometimes lead to false-positive amplification due to the very high specificity and sensitivity (Zhou et al, 2014). These results indicate the possibility of the application of LAMP assays in field conditions and resourcepoor laboratories.…”

Section: Discussionmentioning

confidence: 99%

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Loop-mediated isothermal amplification for rapid molecular detection of Enterocytozoon bieneusi in faecal specimens

Nasarudin

Zainudin

Bernadus

et al. 2015

Journal of Medical Microbiology

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A loop-mediated isothermal amplification (LAMP) assay was developed to detect Enterocytozoon bieneusi DNA for the first time from human faecal specimens. Four primers specific for Enterocytozoon bieneusi were designed corresponding to small subunit rRNA gene sequences and tested on 100 human faecal specimens. Thirty-nine of the faecal specimens (39 %) were confirmed positive for Enterocytozoon bieneusi by LAMP compared with 33 % by PCR and 32 % by light microscopy. LAMP yielded 94 % sensitivity and 88 % specificity compared with microscopy (sensitivity 48 %, specificity 76 %). No significant differences in positive detection of Enterocytozoon bieneusi were found among the three methods (P.0.05). However, LAMP has shown a substantial agreement with PCR (k50.78) and fair agreement was demonstrated between microscopy and PCR (k50.25). In conclusion, the LAMP assay proved to be useful as a simplified, rapid, sensitive and specific alternative molecular screening tool in the diagnosis of Enterocytozoon bieneusi in faecal specimens.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Loop-mediated isothermal amplification for rapid molecular detection of Enterocytozoon bieneusi in faecal specimens

Nasarudin

Zainudin

Bernadus

et al. 2015

Journal of Medical Microbiology

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“…A recent study from Bangladesh has shown encouraging results with LAMP with a diagnostic sensitivity of 90.7% and 100% specificity [19]. The laboratory diagnosis of VL has seen tremendous progress during the last decade and will definitely play an important role in VL elimination program by developing more rapid, sensitive, less expensive and non-invasive diagnostic procedures.…”

mentioning

confidence: 99%

Advances in the Diagnosis of Visceral Leishmaniasis

Prajapati¹,

Mehrotra²

2013

J Mol Biomark Diagn

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Visceral Leishmaniasis (VL or Kala azar) is a disease of poor and neglected populations; it affects 79 countries of the world and accounts 58, 000 new cases to each year [1]. Indian subcontinent covers 90% VL cases of the world, among which Bihar state accounts for most cases [2]. In absence of antileishmanial vaccines early and accurate diagnosis holds the key for the control of VL. Till date parasitological method remains the gold standard for diagnosing VL. Microscopic demonstration of amastigotes in splenic aspirates, peripheral blood mononuclear cells and buffy coat has been shown to possess high sensitivities and absolute specificity [3,4]. Serology based diagnostic methods to detect antibodies against different recombinant Leishmania antigens (rK39, rK28, rK16) are also routinely employed to detect active cases in the field conditions. Out of these rK39 immunochromatographic test (ICT) remains the test of choice for clinicians because of its 100% sensitivity in VL subjects and 85-100% specificity in endemic healthy controls [5]. Recently recombinant antigen rK16, 39-amino acid protein obtained from C terminus of L. donovani immobilized in two different formats, a flow through test (KEFT) or Signal KA, and a lateral flow test (KELF) or Crystal KA had shown higher (sensitivity 93-99% and 99.5% respectively) in Indian subcontinent than East Africa and Brazilian VL subjects [6,7]. rK28 was designed to increase antigen epitope density from L. donovani haspb1 and rK39 kinesin gene. In ELISA, it has shown 100% sensitivity in VL subjects and 94% specificity in endemic healthy controls [6]. Antibody based detection methods have two major limitations. Although antibody titre level goes down after successful VL treatment, it still can be detected after many years. Secondly, house hold contacts from VL patient families have shown positivity for antileishmanial antibodies. Sero-prevalence in these individuals varies from 10-30% and they are termed as asymptomatic cases. Therefore, antibody based approaches should be confirmed with other VL defining tools before treatment [8]. In order to develop non invasive diagnosis for VL, the strip tests have been evaluated using human saliva which is a good source of anti-leishmanial antibodies. Sensitivity for rK39 antigen using saliva was 83.3% and 82.5% in ELISA and ICT technique, whereas specificity was found to be 90.5% and 91.5% respectively by ELISA and ICT in endemic healthy controls [9]. A recent study from Bangladesh has shown satisfactory results with urine samples of VL patients and endemic controls with rK39 based ICT (93.3% specificity and 95% sensitivity) [10]. Since the reliability of these RDTs in any region depends upon the manufacturer, therefore quality and performance should be checked before implementing these RDTs for VL diagnostic algorithm. In immune-compromised patients (HIV-VL co-infection) antibody titre is low therefore RDTs should be evaluated properly in clinical settings to define VL subjects and treatment recommendations.Antigen based diagnosis is...

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