Detecting Cryptosporidium parvum and Giardia lamblia by coagulation concentration and real-time PCR quantification

2012 ◽  
Vol 7 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Huining Zhang ◽  
Xiaohu Zhang ◽  
Shuting Zhang ◽  
Bo Wei ◽  
Qipei Jiang ◽  
...  
Keyword(s):  
Real Time ◽  
Cryptosporidium Parvum ◽  
Real Time Pcr ◽  
Giardia Lamblia

Detection of Cryptosporidium parvum and Giardia lamblia in Water Supply by Quantitative Real-Time PCR

2012 ◽  
Vol 518-523 ◽  
pp. 3707-3711
Author(s):  
Peng Peng ◽  
Rui Bao Jia ◽  
Yu Mei Liu ◽  
Li Li
Keyword(s):  
Water Supply ◽  
Real Time ◽  
Cryptosporidium Parvum ◽  
Real Time Pcr ◽  
Giardia Lamblia ◽  
Pcr Primers ◽  
Quantitative Real Time Pcr ◽  
Rt Pcr ◽  
High Recovery ◽  
Pcr Method

Cryptosporidium parvum and Giardia lamblia are common pathogenic protozoa in water, which pose high risk to drinking water supply. In the present study, detection of C. parvum and G. lamblia was performed by quantitative real-time PCR (RT-PCR). Pairs of PCR primers were evaluated for the detection specificity to pathogenic C. parvum and G. lamblia. The recovery of the RT-PCR detection procedure was examined and high recovery rates (i.e., more than 45％ for C. parvum and more than 50％ for G. lamblia ) were achieved. The RT-PCR method was used to detect C. parvum and G. lamblia in a secondary water supply. The results indicated the potential application of the quantitative RT- PCR method in detection of C. parvum and G. lamblia in water supply.

scholarly journals Genotype-specific detection of Giardia lamblia in stool samples of diarrhoeal and non-diarrhoeal patients in Dhaka, Bangladesh

1970 ◽  
Vol 20 (2) ◽  
pp. 183-189 ◽  
Author(s):  
Md Masud Alam ◽  
Mohammad Ilias ◽  
Md Abdullah Siddique ◽  
Md Mamun Kabir ◽  
Farida Nazib ◽  
...  
Keyword(s):  
Positive Result ◽  
Real Time ◽  
Real Time Pcr ◽  
Giardia Lamblia ◽  
Specific Antigen ◽  
Diarrhoeal Disease ◽  
Giardia Cyst ◽  
Positive Stool ◽  
Stool Samples ◽  
Assemblage B

Two major genotypic assemblages (A and B) of Giardia lamblia infect humans. A single-vessel multiplex real-time PCR assay was used that genotypes Giardia infections into assemblages A and/or B directly from fecal samples. In this study, 157 diarrhoeal (symptomatic) and non-diarrhoeal (asymptomatic) stool samples collected from the International Centre for Diarrhoeal Disease Research, Bangladesh (ICDDR,B) and Bangabandhu Sheikh Mujib Medical University (BSMMU) hospital, respectively were analyzed to determine whether an association exists between infections with G. lamblia assemblages A or B and diarrhea in Bangladesh. Of the 157 stool samples, Giardia cysts were observed in 35 by microscopy and 127 showed positive result for Giardia cyst specific antigen. The 127 ELISA positive samples were assayed for genotyping by real?time polymerase chain reaction. Of the 117 real-time PCR positive stool samples, 15 were positive for G. lamblia assemblage A, 96 were positive for assemblage B and 6 samples showed positive result for both G. lamblia assemblage A and B infections. Higher ratios for diarrhea were observed for assemblage A infections, whereas higher parasite DNA loads and a higher overall rate were observed for assemblage B infections in both diarrhoeal and non-diarrhoeal patients. Real-time PCR is, therefore, useful as an additional test supplementary to microscopy or enzyme immunoassay to detect genotypes of Giardia. Key words: Giardia lamblia; Genotypes; Multiplex real-time PCR; Immunoassay DOI: http://dx.doi.org/10.3329/dujbs.v20i2.8979 DUJBS 2011; 20(2): 183-189

Improved specificity for Giardia lamblia cyst quantification in wastewater by development of a real-time PCR method

2004 ◽  
Vol 57 (1) ◽  
pp. 41-53 ◽  
Author(s):  
Isabelle Bertrand ◽  
Christophe Gantzer ◽  
Thierry Chesnot ◽  
Janine Schwartzbrod
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Giardia Lamblia ◽  
Pcr Method

Quantification of Cryptosporidium parvum in anaerobic digesters treating manure by (reverse-transcription) quantitative real-time PCR, infectivity and excystation tests

2006 ◽  
Vol 53 (8) ◽  
pp. 195-202 ◽  
Author(s):  
G. Garcés ◽  
M. Effenberger ◽  
M. Najdrowski ◽  
C. Wackwitz ◽  
A. Gronauer ◽  
...  
Keyword(s):  
Anaerobic Digestion ◽  
Real Time ◽  
Cryptosporidium Parvum ◽  
Reverse Transcription ◽  
Real Time Pcr ◽  
Combined Treatment ◽  
Quantitative Real Time Pcr ◽  
Anaerobic Digesters ◽  
Hsp70 Mrna ◽  
Cryptosporidium Parvum Oocysts

The survival of Cryptosporidium parvum oocysts in anaerobic digesters treating manure was investigated for mesophilic, thermophilic, and a combined treatment (mesophilic–thermophilic–mesophilic) under different retention times of oocysts in the reactors. C. parvum DNA was extracted with an optimised protocol, and its amount determined by quantitative real-time PCR (qPCR). Results indicated noteworthy differences in DNA content after the different treatments. DNA was not degraded during the process. However, excystation and infectivity tests showed a reduction of viable oocyst numbers of ≥2 and ≥5 log units after the thermophilic treatment in two different experiments. Thus qPCR-targeting DNA can overestimate the number of oocysts that survive and remain viable after anaerobic digestion. However, targeting DNA is suitable to indicate the presence or absence of oocysts. Reverse transcription qPCR (RT-qPCR) targeting C. parvum hsp70 mRNA successfully indicated the presence of viable fraction of oocysts.

Evaluation of alternative DNA extraction processes and real-time PCR for detecting Cryptosporidium parvum in drinking water

2015 ◽  
Vol 15 (6) ◽  
pp. 1295-1303
Author(s):  
Gina H. Kimble ◽  
Vincent R. Hill ◽  
James E. Amburgey
Keyword(s):  
Drinking Water ◽  
Real Time ◽  
Dna Extraction ◽  
Cryptosporidium Parvum ◽  
Water Samples ◽  
Real Time Pcr ◽  
Extraction Procedure ◽  
The United States ◽  
Detection Sensitivity ◽  
Spin Column

USEPA Method 1623 is the standard method in the United States for the detection of Cryptosporidium in water samples, but quantitative real-time polymerase chain reaction (qPCR) is an alternative technique that has been successfully used to detect Cryptosporidium in aqueous matrices. This study examined various modifications to a commercial nucleic acid extraction procedure in order to enhance PCR detection sensitivity for Cryptosporidium. An alternative DNA extraction buffer allowed for qPCR detection at lower seed levels than a commercial extraction kit buffer. In addition, the use of a second spin column cycle produced significantly better detection (P = 0.031), and the volume of Tris–EDTA buffer significantly affected crossing threshold values (P = 0.001). The improved extraction procedure was evaluated using 10 L of tap water samples processed by ultrafiltration, centrifugation and immunomagnetic separation. Mean recovery for the sample processing method was determined to be 41% using microscopy and 49% by real-time PCR (P = 0.013). The results of this study demonstrate that real-time PCR can be an effective alternative for detecting and quantifying Cryptosporidium parvum in drinking water samples.

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