The simultaneous detection of both enteroviruses and adenoviruses in environmental water samples including tap water with an integrated cell culture-multiplex-nested PCR procedure

2005 ◽  
Vol 98 (5) ◽  
pp. 1020-1029 ◽  
Author(s):  
S.-H. Lee ◽  
C. Lee ◽  
K.W. Lee ◽  
H.B. Cho ◽  
S.-J. Kim
Keyword(s):  
Cell Culture ◽  
Water Samples ◽  
Nested Pcr ◽  
Tap Water ◽  
Simultaneous Detection ◽  
Environmental Water ◽  
Environmental Water Samples

Rotavirus detection in environmental water samples by tangential flow ultrafiltration and RT-nested PCR

2009 ◽  
Vol 164 (1-4) ◽  
pp. 199-205 ◽  
Author(s):  
Tiziana Grassi ◽  
Francesco Bagordo ◽  
Adele Idolo ◽  
Federica Lugoli ◽  
Giovanni Gabutti ◽  
...  
Keyword(s):  
Water Samples ◽  
Nested Pcr ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Tangential Flow

An efficient virus concentration method and RT-nested PCR for detection of rotaviruses in environmental water samples

2005 ◽  
Vol 124 (1-2) ◽  
pp. 117-122 ◽  
Author(s):  
Leera Kittigul ◽  
Som Ekchaloemkiet ◽  
Fuangfa Utrarachkij ◽  
Kanokrat Siripanichgon ◽  
Dusit Sujirarat ◽  
...  
Keyword(s):  
Water Samples ◽  
Nested Pcr ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Virus Concentration ◽  
Concentration Method ◽  
Virus Concentration Method

scholarly journals Real-Time PCR for Quantification of Giardia and Cryptosporidium in Environmental Water Samples and Sewage

2003 ◽  
Vol 69 (9) ◽  
pp. 5178-5185 ◽  
Author(s):  
Rebecca A. Guy ◽  
Pierre Payment ◽  
Ulrich J. Krull ◽  
Paul A. Horgen
Keyword(s):  
Real Time ◽  
Water Samples ◽  
Real Time Pcr ◽  
Simultaneous Detection ◽  
Environmental Water ◽  
Qpcr Assay ◽  
Detection Methods ◽  
Environmental Water Samples ◽  
Dna Yield ◽  
Chelex 100

ABSTRACT The protozoan pathogens Giardia lamblia and Cryptosporidium parvum are major causes of waterborne enteric disease throughout the world. Improved detection methods that are very sensitive and rapid are urgently needed. This is especially the case for analysis of environmental water samples in which the densities of Giardia and Cryptosporidium are very low. Primers and TaqMan probes based on the β-giardin gene of G. lamblia and the COWP gene of C. parvum were developed and used to detect DNA concentrations over a range of 7 orders of magnitude. It was possible to detect DNA to the equivalent of a single cyst of G. lamblia and one oocyst of C. parvum. A multiplex real-time PCR (qPCR) assay for simultaneous detection of G. lamblia and C. parvum resulted in comparable levels of detection. Comparison of DNA extraction methodologies to maximize DNA yield from cysts and oocysts determined that a combination of freeze-thaw, sonication, and purification using the DNeasy kit (Qiagen) provided a highly efficient method. Sampling of four environmental water bodies revealed variation in qPCR inhibitors in 2-liter concentrates. A methodology for dealing with qPCR inhibitors that involved the use of Chelex 100 and PVP 360 was developed. It was possible to detect and quantify G. lamblia in sewage using qPCR when applying the procedure for extraction of DNA from 1-liter sewage samples. Numbers obtained from the qPCR assay were comparable to those obtained with immunofluorescence microscopy. The qPCR analysis revealed both assemblage A and assemblage B genotypes of G. lamblia in the sewage. No Cryptosporidium was detected in these samples by either method.

scholarly journals Simultaneous direct detection of toxigenic and non-toxigenic Vibrio cholerae from rectal swabs and environmental samples by sandwich ELISA

2007 ◽  
Vol 56 (10) ◽  
pp. 1340-1345 ◽  
Author(s):  
Urmil Tuteja ◽  
Sanjay Kumar ◽  
Jyoti Shukla ◽  
Joseph Kingston ◽  
Harsh V. Batra
Keyword(s):  
Vibrio Cholerae ◽  
Water Samples ◽  
Direct Detection ◽  
Polyclonal Antibodies ◽  
Sandwich Elisa ◽  
Simultaneous Detection ◽  
Environmental Water ◽  
Environmental Water Samples ◽  
Bacterial Strains ◽  
Rectal Swabs

A mAb-based simple, specific and rapid two-tip dipstick ELISA was developed for simultaneous detection of toxin- and non-toxin-producing strains of Vibrio cholerae, and for direct detection of V. cholerae from rectal swabs of patients and from environmental water samples. Rabbit polyclonal antibodies and murine mAbs were raised against recombinant protein (r-protein) antigens of cholera toxin B (CtxB) and outer membrane protein W (OmpW). Rabbit polyclonal antibodies to both r-proteins were coated individually onto the tips of nitrocellulose (NC) membranes of a two-tipped NC dipstick as capture antibodies and a mixture of two mAbs was used for the detecting antibodies. The test was found to be specific for V. cholerae strains O1, O139, non-O1 and non-O139, and did not show any cross-reaction to closely related bacterial strains. The test was evaluated on rectal swabs collected at the bedside of 75 hospitalized diarrhoeal patients and on 50 environmental water samples after enrichment for 4 h in alkaline peptone water. The mAb two-tip dipstick ELISA detected V. cholerae in 52/75 rectal swabs and 2/50 environmental water samples for CtxB antigen, and in 1/50 environmental water samples for the non-toxin OmpW antigen of V. cholerae within 1.5 h. These findings were identical to those observed using PCR and conventional culture methods. Thus, this mAb-based two-tip dipstick ELISA could be used for early and reliable simultaneous detection of toxigenic and non-toxigenic strains of V. cholerae from clinical and environmental water samples.

scholarly journals Determination of OCPs and PCBs in environmental water samples by GC-DLLME optimized by Response Surface Methodology

2019 ◽  
Vol 20 (46) ◽  
pp. 13-23
Author(s):  
Dekun Hou ◽  
Otgonbayar Khureldavaa ◽  
Fujin Zhang ◽  
Jiang He ◽  
Badgaa Amarsanaa
Keyword(s):  
Response Surface Methodology ◽  
Response Surface ◽  
Lake Water ◽  
Water Samples ◽  
Tap Water ◽  
Environmental Water ◽  
Enrichment Factors ◽  
Environmental Water Samples ◽  
Relative Standard

A new sample preparation procedure to determine seven organochlorine pesticides and seven polychlorinated biphenyls in environmental water samples by using a combination of ultrasonic-assisted solvent extraction and dispersive liquid-liquid micro-extraction was established, and the extracted analytes were analyzed by gas chromatography coupled with electron capture detector. Some parameters influencing the extraction efficiency were studied and optimized utilizing response surface methodology. Under the optimum extraction conditions, the method showed wide linear ranges with r2 > 0.9989 and low limits of detection and quantification between 0.16 ~ 2.17 μg/L and 0.53 ~ 7.16 μg/L, respectively. Enrichment factors (EF) were high and ranged from 63 to 116. Relative standard deviations (RSDs) for the extraction of 25 μg/L of each selected OCPs and PCBs were less than 10.2 %. The proposed method was successfully used for targets contaminations determination in different water samples. α-HCH, β-HCH and p,p’-DDE were found in lake water closed to farmland with concentrations of 2.56 μg/L, 4.44 μg/L and 4.74 μg/L, respectively, and other OCPs and PCBs were not found in the corresponding water samples. The relative recoveries of OCPs and PCBs from tap water, river water and lake water at spiking levels of 10 μg/L were in the range of 81.9 ~ 109.7 %, within a relative standard deviation of 1.7 ~ 11.8 %. The results revealed that the proposed method was well suited for the determination of trace amounts of target contaminations in liquid samples.

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