Development and Preliminary Evaluation of a Loop-Mediated Isothermal Amplification Procedure for Sensitive Detection of Cryptosporidium Oocysts in Fecal and Water Samples

Abstract

A loop-mediated isothermal amplification (LAMP) procedure for the detection of Cryptosporidium in environmental and fecal samples was developed and evaluated. This is the first demonstration of LAMP applied to detection of Cryptosporidium. Due to its specificity and simplicity, the method could become a useful diagnostic tool for epidemiologic studies of Cryptosporidium presence.

Detection of Early and Single Infections of Schistosoma japonicum in the Intermediate Host Snail, Oncomelania hupensis, by PCR and Loop-Mediated Isothermal Amplification (LAMP) Assay

“…Our results showed that the sensitivity of LAMP was the same as that of conventional PCR. In general, the LAMP method is more sensitive than the PCR method, 20,22,34,35 although similar sensitivities between the two have also been reported. 24 This may reflect the fact that the sensitivity is dependent on the designed primers.…”

Theileria annulata DNA’sının loop aracılı izotermal yöntemle (LAMP) çoğaltılması

“…LAMP yöntemi son yıllarda, tek nükleotid polimorfizmlerinin belirlenmesine dayalı genetik çalışmalar ile viral, bakteriyel ve paraziter etkenlerin tanısal amaçlı belirlenmesinde giderek artan oranda kullanılmaya başlan-mıştır. Paraziter kökenli hastalıklardan equine piroplasmosis (2,3,32), tropikal theileriosis (15,28), East Cost Fever (ECF) (29) ile sığır babesiosis (10,14) etkenlerinin ve koyunlarda hastalık oluşturan Theileria ve Babesia soyundaki kan parazitlerinin (8,16,27), Giardia duodenalis (26) ve Cryptosporidium türlerinin (5,12), Trypanosoma brucei gambiense'nin (13), köpeklerde B.gibsoni (17,9) ve B.canis (1,19) ile ara konak sivrisineklerdeki Plasmodium spp. 'lerin (4) teşhisi amacıyla LAMP yön-temi kullanılarak birçok çalışma yapılmıştır.…”

Advances in Diagnosis of Important Protozoan Diseases: Old and New Approaches

“…Recently this technique has been adapted to detect African trypanosomes. A number of primers have been described for the detection of T. brucei, T. congolense, T. vivax, Trypanosoma gambiense and T. evansi (Kuboki et al,2003, Njiru et al, 2008and Thekisoe et al,2007.…”

“…Real-time PCR detection of protozoan parasites (Table 1) LAMP Alternative DNA amplification, such as loop mediated isothermal amplification (LAMP) has been applied to protozoa (Alhassan et al, 2007, Karanis et al, 2007, Njiru et al, 2008, Guan et al, 2008and Karanis and Ongerth, 2009. In this method, six different primers, specifically designed to recognise eight distinct regions on a target gene, with amplification only occurring if all the primers bind and form a product.…”