1887

Abstract

Loop-mediated isothermal amplification (LAMP) is a novel, rapid nucleic acid amplification method with high specificity and sensitivity under isothermal conditions. In this study a LAMP assay for diagnosing pneumonia (PCP) was developed. Oligonucleotide primers specific for species were designed corresponding to 18S rRNA gene sequences. The assay, performed for 30 min at 61 °C, was capable of detecting 50 copies per tube (2×10 copies ml) in 30 min and did not show cross-reactivity to other species of fungi, including the genera , and . A total of 21 of 24 clinical specimens (sputum and bronchoalveolar lavage fluid) from patients with suspected PCP tested positive using the LAMP assay by real-time fluorescence detection. The results of the LAMP reaction were also observed by real-time turbidity detection and end-point visual turbidity or fluorescence detection. With real-time fluorescence detection, melting curves of the products were effective at distinguishing specific amplification from non-specific amplification or self-amplification. Visual detection was also possible as a rapid and easy assay using only a heat block and a black light.

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2008-01-01
2021-10-28
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