Detection of Acanthamoeba in Tap Water and Contact Lens Cases Using Polymerase Chain Reaction

2008 ◽  
Vol 85 (7) ◽  
pp. 526-530 ◽  
Author(s):  
Maureen Boost ◽  
Pauline Cho ◽  
Sindy Lai ◽  
Wing-Man Sun
Keyword(s):  
Polymerase Chain Reaction ◽  
Contact Lens ◽  
Tap Water ◽  
Chain Reaction ◽  
Polymerase Chain

Detection of small round structured viruses in clinical and environmental samples by polymerase chain reaction

1995 ◽  
Vol 31 (5-6) ◽  
pp. 375-382 ◽  
Author(s):  
M. Wolfaardt ◽  
C. L. Moe ◽  
W. O. K. Grabow
Keyword(s):  
Polymerase Chain Reaction ◽  
Tap Water ◽  
Practical Method ◽  
Glass Wool ◽  
Polluted Water ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Elution Method

Norwalk (NV) and other small round structured viruses (SRSVs) have been identified as common causes of gastroenteritis. Outbreaks of Norwalk gastroenteritis have been associated with contaminated drinking water and food such as oysters and salads. The cloning and sequencing of the NV genome has made it possible to detect NV and related viruses by the reverse transcription-polymerase chain reaction (RT-PCR). We applied RT-PCR to detect SRSVs in faecal specimens from two gastroenteritis outbreaks in South Africa, designated “Christmas” and “Grootbrak” and were able to detect SRSVs in all of the three specimens from the Christmas outbreak and in two of 16 specimens from the Grootbrak outbreak. The RT-PCR procedure used appeared to be more sensitive for the detection of SRSVs in patient stool specimens than immune electron microscopy and NV antigen detection by enzyme linked immunosorbent assay. The RT-PCR procedure proved suitable for the detection of SRSVs in seeded samples of sewage, sewage sludge, river water, and tap water. However, sensitivity was lower for seeded samples of sewage and sludge than for tap water, which indicates interference by high levels of organic matter. The RT-PCR procedure was also used to show that small numbers of SRSVs can successfully be recovered from large volumes of water by means of a glass wool adsorption-elution method. Since no practical method is available for quantitation of the small numbers of SRSVs concerned, it was not possible to evaluate the efficiency of recovery. Although no SRSVs have been detected by direct testing of sewage and sludge samples, the results obtained in this study show that RT-PCR detection of SRSVs in sewage and polluted water environments is feasible, and that small numbers of the viruses can, like many other enteric viruses, successfully be recovered by means of a glass wool adsorption-elution method.

Development of a Polymerase Chain Reaction Assay for Detection of Burkholderia mallei, a Potent Biological Warfare Agent

2016 ◽  
Vol 66 (5) ◽  
pp. 458 ◽  
Author(s):  
Vijai Pal ◽  
Sandeep Singh ◽  
Arvind Kumar Tiwari ◽  
Y.K. Jaiswal ◽  
G.P. Rai
Keyword(s):  
Polymerase Chain Reaction ◽  
Tap Water ◽  
False Negative ◽  
Burkholderia Mallei ◽  
Chain Reaction ◽  
Pcr Assay ◽  
Negative Results ◽  
False Negative Results ◽  
Polymerase Chain ◽  
Rapid Tool

Burkholderia mallei is the etiological agent of glanders, primarily a disease of equines. B. mallei is closely related to B. pseudomallei, the causative agent of melioidosis. Therefore, detection of B. mallei and its differentiation from B. pseudomallei, has always been troublesome. In present investigation, a B. mallei specific DNA sequence was identified by performing BLASTn search using ~3000 ORFs of B. mallei NCTC 10229. A polymerase chain reaction (PCR) assay with internal amplification control (IAC) was developed for detection of B. mallei and its differentiation from B. pseudomallei. The PCR assay could amplify a specific 224-bp fragment from all the six B. mallei strains used in the study, whereas other closely related organisms were tested negative. The detection limit of the assay was found to be 10 pg of purified DNA of B. mallei. Incorporation of IAC in the assay makes the results reliable as false negative results which may arise due to presence of PCR inhibitors, can be avoided. For validation, the assay was tested on tap water, Bengal gram and grass artificially spiked with B. mallei. The developed assay can be used as a simple and rapid tool for detection of B. mallei.

Effect of Feeding Honey Bee (Apis mellifera Hymenoptera: Apidae) Colonies With Honey, Sugar Solution, Inverted Sugar, and Wheat Starch Syrup on Nosematosis Prevalence and Intensity

2019 ◽  
Author(s):  
Ivana Papežíková ◽  
Miroslava Palíková ◽  
Eva Syrová ◽  
Anna Zachová ◽  
Kristina Somerlíková ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Tap Water ◽  
Wheat Starch ◽  
Sugar Solution ◽  
Nosema Apis ◽  
Chain Reaction ◽  
Bee Colony ◽  
Fitness Parameters ◽  
Polymerase Chain ◽  
Effect Of Feeding

Abstract Here, we present the results of a 2-year field trial aimed at testing the effect of overwintering on different feeds on the course of Nosema ceranae infection. In August 2015, four experimental bee colony groups were established. After the last honey harvest, each colony was provided with 20 kg of feed, either honey, sugar (3:2 solution in tap water), inverted syrup made of sucrose, or wheat starch syrup. Samples of live bees were collected from each beehive in August (before feeding), November, and May. The following year, feeding and sampling were performed in the same way. Bees were examined microscopically to estimate the percentage of Nosema-infected individuals in the sample and the spore number per bee. Fitness parameters were also measured in all colonies. In all hives, presence of N. ceranae was confirmed through polymerase chain reaction. Nosema apis was not detected in the apiary. Significant differences in nosematosis prevalence and/or intensity were observed between the experimental groups. For most parameters, best results were recorded in the group fed with honey. Worst fitness and highest nosematosis prevalence and intensity were found in colonies fed with wheat starch syrup.

Outbreak ofBurkholderia CepaciaBacteremia Traced to Contaminated Hospital Water Used for Dilution of an Alcohol Skin Antiseptic

2004 ◽  
Vol 25 (3) ◽  
pp. 231-239 ◽  
Author(s):  
Rana M. Nasser ◽  
Amal C. Rahi ◽  
Mona F. Haddad ◽  
Ziad Daoud ◽  
Noha Irani-Hakime ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment ◽  
Fragment Length ◽  
Burkholderia Cepacia ◽  
Tap Water ◽  
Bloodstream Infections ◽  
University Hospital ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

AbstractObjective:To identify the source of an epidemic ofBurkholderia cepaciabloodstream infections during 7 years (411 episodes in 361 patients).Design:Outbreak investigation.Setting:A 250-bed university hospital in Beirut, Lebanon.Methods:Matched case-control and retrospective cohort studies, and microbiological surveillance and polymerase chain reaction–restriction fragment length ascertainment were employed. Special media and filtration techniques were used to isolate organisms from water and diluted alcohol solutions.Results:In a group of 50 randomly selected case-matched patients from 1999, the positive blood cultures were concomitant with fever in 98%, intravenous phlebitis in 44%, and recurrent bacteremia in 20%. Fever disappeared approximately 6 hours after intravenous catheter removal. Polymerase chain reaction–restriction fragment length polymorphism revealed strain homogeneity in patient, water, and alcohol isolates. Contaminated tap water had been used to dilute alcohol for skin antisepsis and for decontamination of the caps of heparin vials. Only sporadic cases directly attributable to breach of protocol were reported after single-use alcohol swabs were substituted.Conclusion:This is potentially the largest single-source nosocomial bloodstream infection outbreak ever reported, and the first report of an alcohol skin antiseptic contaminated by tap water as a source for nosocomial bacteremia.

scholarly journals Differences of Legionella Pneumophila Serogroups Distribution in Well Water, Tap Water, Ice Cubes, Hospital and Hotel Water in East Java

2018 ◽  
Author(s):  
Eduardus Bimo Aksono ◽  
Kadek Rachmawati ◽  
Retno Bijanti
Keyword(s):  
Polymerase Chain Reaction ◽  
Phylogenetic Tree ◽  
Legionella Pneumophila ◽  
Water Samples ◽  
Tap Water ◽  
Natural Habitat ◽  
Well Water ◽  
Chain Reaction ◽  
Water Ice ◽  
Polymerase Chain

Background Legionella pneumophila is one of the causes of legionellosis. Water environments serve as the natural habitat and the main sources of Legionella pneumophila. Objectives The aims of this study was to understand the differences of Legionella pneumophila serogroups distribution in well water, tap water, ice cubes, hospital and hotel water in East Java-Indonesia. Methods a total of 60 water samples in east java-Indonesia; from well water (n=25), tap water (n=5), ice cubes (n=5), water from the hospital (n=16), and hotel water (n=9) were detected using polymerase chain reaction with mip gene spesific primers and then it was analyzed by phylogenetic tree. Results For the 60 water samples collected in East Java, 12% of the samples (7/60) were positively contaminated by L. pneumophila. In details, there was 8% of the well water samples (2/25), 2% of the tap water samples (1/5), 2% of the ice cubes samples (1/5), 0% of the hospital water samples (0/16) and 33.33% of the hotel water samples (3/9). The phylogenetic tree showed that Legionella pneumophila contaminating well water isolate 1 from Surabaya and tap water isolate from Sidoarjo was closer to L.pneumophila serogroup 2, 4, 6, 8, 10, 12, isolates from Brazil, China, Spain and Australia. L.pneumophila contaminating the ice cubes from Sidoarjo was closer to serogroup 1, 2, 4, 7, 8, 11, 13, 14, while the bacteria contaminating well water isolate 2 from Sidoarjo as well as water in hotel of Surabaya (hotel water isolate 1, 2 and 3) classified into their own group. Conclusion There is a difference in the distribution of L. pneumophila serogroups between well water, tap water, ice cubes, and hotels.

High performance Nanogold™-in situ hybridzation and its use in the detection of hybridized and PCR-amplified microscopical preparations

1996 ◽  
Vol 54 ◽  
pp. 896-897
Author(s):  
G. W. Hacker ◽  
I. Zehbe ◽  
J. Hainfeld ◽  
A.-H. Graf ◽  
C. Hauser-Kronberger ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Messenger Rna ◽  
High Performance ◽  
Detection System ◽  
Direct Methods ◽  
Genetic Alterations ◽  
Chain Reaction ◽  
Protein Encoding ◽  
Polymerase Chain

In situ hybridization (ISH) with biotin-labeled probes is increasingly used in histology, histopathology and molecular biology, to detect genetic nucleic acid sequences of interest, such as viruses, genetic alterations and peptide-/protein-encoding messenger RNA (mRNA). In situ polymerase chain reaction (PCR) (PCR in situ hybridization = PISH) and the new in situ self-sustained sequence replication-based amplification (3SR) method even allow the detection of single copies of DNA or RNA in cytological and histological material. However, there is a number of considerable problems with the in situ PCR methods available today: False positives due to mis-priming of DNA breakdown products contained in several types of cells causing non-specific incorporation of label in direct methods, and re-diffusion artefacts of amplicons into previously negative cells have been observed. To avoid these problems, super-sensitive ISH procedures can be used, and it is well known that the sensitivity and outcome of these methods partially depend on the detection system used.

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