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J Med Microbiol. 2003 Aug;52(Pt 8):681-3.

Detection and genotyping by real-time PCR/RFLP analyses of Giardia duodenalis from human faeces.

Author information

1
Health Protection Agency, Food Safety Microbiology Laboratory, Division of Gastrointestinal Infections, Central Public Health Laboratory, 61 Colindale Avenue, London NW9 5HT, UK.

Abstract

A nested PCR assay (TPILC-PCR) was developed to detect and distinguish between Giardia duodenalis assemblages A and B from human faeces by analysis of the triose phosphate isomerase gene (tpi). The assay comprised an initial multiplexed block-based amplification. This was followed by two separate real-time PCR assays specific for assemblages A and B using a LightCycler and SYBR Green I to identify PCR products by melting-point analysis. RFLP analysis was applied to distinguish G. duodenalis assemblage A groups I and II. The real-time nested PCR was evaluated using DNA extracted from purified giardial trophozoites, Cryptosporidium oocysts, whole faeces containing a range of potential pathogens (including G. duodenalis), faecal smears and bacterial suspensions. The assay was specific, sensitive, reproducible and rapid.

PMID:
12867562
DOI:
10.1099/jmm.0.05193-0
[Indexed for MEDLINE]

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