Specific, Sensitive, and Rapid Diagnosis of Active Toxoplasmosis by a Loop-Mediated Isothermal Amplification Method Using Blood Samples from Patients

Lau, Yee Ling; Meganathan, Puviarasi; Parthasarathy, S.; Thiruvengadam, Girija; Nissapatorn, Veeranoot; Chen, Yeng

doi:10.1128/jcm.00462-10

Cited by 90 publications

(71 citation statements)

References 31 publications

(25 reference statements)

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“…Several reports were confirmed the higher sensitivity of LAMP for detection of T. gondii than conventional PCR targeting 529 bp fragment of T. gondii genome (Fallahi et al 2014;Kong et al 2012;Lin et al 2012;Zhang et al 2009) or other specific targeting of T. gondii DNA regions (Fallahi et al 2014;Hu et al 2012;Kong et al 2012;Krasteva et al 2009;Lau et al 2010;Lin et al 2012;Sotiriadou and Karanis 2008;Zhang et al 2009). Until now, LAMP method was used for detection of T. gondii in pig (Lili et al 2014;Lin et al 2012;Qu et al 2013;Wang et al 2013;Zhang et al 2009), sheep (Lin et al 2012), mice (Hu et al 2012;Kong et al 2012;Krasteva et al 2009), and human (Fallahi et al 2014;Lau et al 2010). LAMP was also used for detection of T. gondii oocysts in water samples (GallasLindemann et al 2013;Koloren and Demirel 2013;Sotiriadou and Karanis 2008).…”

Section: Discussionmentioning

confidence: 80%

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

et al. 2015

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Toxoplasma gondii is one of the most common zoonotic parasitic diseases in human and warm-blooded animals worldwide. Birds are one of important intermediate hosts of T. gondii. The aim of this study is molecular detection of T. gondii in the house sparrow by LAMP and PCR methods in Tehran, Iran. A total 200 sparrows were captured in different regions of Tehran. DNA was extracted from tissue samples of each sparrow. LAMP and conventional PCR assays were carried out with a set of primers to detect the 529 bp fragment of T. gondii. LAMP and PCR were detected T. gondii from 17 (8.5 %) and 15 (7.5 %) of 200 sparrows respectively. These results indicated that sensitivity of LAMP was higher than conventional PCR. In our knowledge, this study is the first report of detection of T. gondii by LAMP method in bird hosts. Also, these findings provided an insight into epidemiological pattern of T. gondii infection in sparrow in Iran.

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Section: Discussionmentioning

confidence: 80%

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

et al. 2015

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“…As a result, this technology has been used widely for molecular detection of several microorganisms, including clinical and plant-associated bacterial [32,[61][62][63][64][65][66][67][68][69][70][71][72][73], fungal [74][75][76], viral [47,[77][78][79][80][81][82][83][84], and parasitic [57,59,[85][86][87][88] pathogens, and is therefore suitable as a rapid field test.…”

Section: Discussionmentioning

confidence: 99%

Specific Detection of Klebsiella variicola and K. oxytoca by Loop-Mediated Isothermal Amplification

Yasuhara-Bell¹,

Ayin²,

Hatada³

et al. 2015

J Plant Pathol Microbiol

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Klebsiella spp. are opportunistic pathogens with clinical, veterinary and plant-associated isolates. A previous study showed that bacterial ooze from wetwood of severely declined ironwood trees in Guam contained Ralstonia solanacearum, Klebsiella variicola and K. oxytoca. In this study, Loop-Mediated Isothermal Amplification (LAMP) detected K. variicola and K. oxytoca specifically, using unique primer sets designed individually for each organism. Each LAMP detected its target specifically, while showing negative results for non-target bacteria and negative controls. LAMP detected Klebsiella in inoculated-ironwood stem tissues and bacterial ooze. Due to the presence of plant inhibitors, different sampling protocols were tested. Soaking plant tissue samples to allow diffusion of bacteria into solution, followed by boiling, provided optimum detection of Klebsiella directly from plant samples. False negatives obtained when using crushed plant samples were eliminated by including an enrichment step, involving plating and 12-h incubation. DGGE (denaturing gradient gel electrophoresis) and a colony-blot immunoassay using a Klebsiella-specific antibody also detected Klebsiella in inoculated ironwood. DGGE bands and antibody crossreactions from closely related enterobacters showed the potential for false positive results. The nature of LAMP makes it ideal for point-of-care testing, and when combined with the specificity of the LAMP primers developed in this study, demonstrates its potential as a routine field test for Klebsiella in ironwood in Guam, as well as clinical and veterinary diagnosis of Klebsiella infection. Additionally, the regions targeted for detection in this study have application across all forms of molecular-based diagnostics.

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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%

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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Specific, Sensitive, and Rapid Diagnosis of Active Toxoplasmosis by a Loop-Mediated Isothermal Amplification Method Using Blood Samples from Patients

Cited by 90 publications

References 31 publications

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

Molecular detection of Toxoplasma gondii in house sparrow (Passer domesticus) by LAMP and PCR methods in Tehran, Iran

Specific Detection of Klebsiella variicola and K. oxytoca by Loop-Mediated Isothermal Amplification

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

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