Real-Time PCR Assay for Detection and Genotype Differentiation of Giardia lamblia in Stool Specimens

A rapid laboratory system has been developed and evaluated that can simultaneously identify major diarrhoeagenic bacteria, including Salmonella enterica, Vibrio parahaemolyticus, Campylobacter jejuni and Shiga toxin-producing Escherichia coli, in stool specimens by real-time PCR. Specific identification was achieved by using selective TaqMan probes, detecting two targets in each pathogen. A positive result was scored only when both targets of a pathogen were amplified and the difference between threshold cycles for detection was less than five. Diagnosis of enteric bacterial infections using this highly sensitive method, including DNA extraction and real-time PCR, requires only 3 h. Forty stool specimens related to suspected food poisoning outbreaks were analysed: 16 (40 %) of these samples were found to be positive for diarrhoeagenic bacteria using a conventional culture method; 28 (70 %) were positive using the real-time PCR assay. Of the 12 PCR-positive but culture-negative cases, 11 patients had consumed pathogen-contaminated or high-risk food. Analysis of faecal samples from 105 outpatients who complained of diarrhoea and/or abdominal pain identified 19 (18 %) patients as being positive for diarrhoeagenic bacteria using the culture method. An additional six (6 %) patients were found to be positive by PCR analysis.

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Evaluation of a new real-time PCR assay for the direct detection of diarrheagenic Escherichia coli in stool specimens

Diagnostic Microbiology and Infectious Disease ◽

10.1016/j.diagmicrobio.2017.01.016 ◽

2017 ◽

Vol 88 (1) ◽

pp. 12-16 ◽

Cited By ~ 6

Author(s):

U. Eigner ◽

A. Hiergeist ◽

A. Veldenzer ◽

M. Rohlfs ◽

R. Schwarz ◽

...

Keyword(s):

Escherichia Coli ◽

Real Time ◽

Real Time Pcr ◽

Direct Detection ◽

Pcr Assay ◽

Diarrheagenic Escherichia Coli ◽

Stool Specimens

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Detection of Giardia lamblia in children by microscopy, ELISA and real time PCR assay

Dhaka University Journal of Biological Sciences ◽

10.3329/dujbs.v23i2.20100 ◽

2014 ◽

Vol 23 (2) ◽

pp. 197-204 ◽

Cited By ~ 1

Author(s):

Tahmina Ahmed ◽

Hamida Khanum ◽

Priyanka Barua ◽

Tuhinur Arju ◽

Muhammed Salah Uddin ◽

...

Keyword(s):

Real Time ◽

Real Time Pcr ◽

Giardia Lamblia ◽

Urban Slum ◽

Pcr Assay ◽

Asymptomatic Children ◽

Faecal Samples ◽

Female Children ◽

Conventional Microscopy

Conventional microscopy, ELISA and Real Time PCR showed Giardia lamblia in a total of 4470 asymptomatic and 423 symptomatic faecal samples from urban slum children of 13 ? 60 months age old. ELISA showed more sensitivity (88.88%) and specificity (90.91%) than microscopy (6.5% sensitive and 100% specific). The prevalence of G. lamblia infection among asymptomatic children was 37.93% by microscopy and 76.11% by ELISA. The prevalence was 7.03, 38.91 and 66.49 by microscopy, ELISA and Real Time PCR in symptomatic children, respectively. The highest prevalence (80.95%) of infection was found in symptomatic children between 49 and 60 months old. Peak prevalence among asymptomatic children was found in March, 2012 (39.39%) and among symptomatic children in September, 2012 (88.88%). Giardia lamblia infection was higher (71.03%) in male than in female children (60.26%). DOI: http://dx.doi.org/10.3329/dujbs.v23i2.20100 Dhaka Univ. J. Biol. Sci. 23(2): 197-204, 2014

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