scholarly journals Comparison of three laboratorial tests for diagnosis of canine parvovirus infection

2013 ◽  
Vol 65 (1) ◽  
pp. 149-152 ◽  
Author(s):  
M.M.O. Silva ◽  
T.X. Castro ◽  
E.M. Costa ◽  
T.A.L. Trancoso ◽  
F. Mendes-de-Almeida ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Enzyme Immunoassay ◽  
Antigen Detection ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Fecal Samples ◽  
Negative Results ◽  
Rapid Tests ◽  
Polymerase Chain ◽  
One Year

The aim of this study was to evaluate the rapid tests currently used for canine parvovirus (CPV) diagnosis: hemagglutination test (HA), enzyme immunoassay (EIA) and polymerase chain reaction (PCR). A total of 112 fecal samples collected from diarrheic puppies up to one year of age were tested. The EIA was able to detect CPV antigen in 44 samples. By HA, 32 samples tested highly positive with titers >128, eight tested weakly positive (titers 32 and 64) and 72 were negative (titers <16). Using PCR, 57 samples were found positive including 13 EIA-negative and 19 HA-negative samples. The best correlation was observed between EIA and PCR (88.4%). These tests were able to detect all types of CPV, including CPV-2c. Considering that 23%-33% of dogs presenting enteritis did not show infection by EIA nor HA, negative results from the antigen detection tests should be confirmed through molecular methods.

scholarly journals The Use of a Mimic to Detect Polymerase Chain Reaction– Inhibitory Factors in Feces Examined for the Presence of Lawsonia Intracellularis

2003 ◽  
Vol 15 (3) ◽  
pp. 268-273 ◽  
Author(s):  
Magdalena Jacobson ◽  
Stina Englund ◽  
András Ballagi-Pordány
Keyword(s):  
Polymerase Chain Reaction ◽  
False Negative ◽  
Clinical Samples ◽  
Lawsonia Intracellularis ◽  
Chain Reaction ◽  
Fecal Samples ◽  
Wild Boars ◽  
Negative Results ◽  
False Negative Results ◽  
Polymerase Chain

Lawsonia intracellularis is an intracellular organism that causes proliferative enteritis in pigs. This bacterium is difficult to culture, and antemortem demonstration of the microbe is therefore often performed on fecal samples by polymerase chain reaction (PCR). Polymerase chain reaction is sensitive and specific, but inhibitory factors in feces might cause false-negative results. This article describes the construction and use of an internal standard, a mimic. The mimic is amplified by the same primers as those used for L. intracellularis DNA and thus could indicate false-negative results in clinical samples. The amplicon was clearly visible when as few as 10 mimic molecules were added per amplification reaction and when no inhibitors were present. When fecal samples were spiked with the mimic, the detection limit was 102 molecules per PCR. Sixty clinical samples, 20 from wild boars, 20 from growing pigs with diarrhea, and 20 from pigs without diarrhea, were prepared by a boiling procedure and subjected to PCR together with 103 mimic molecules. Nine samples were positive, of which 7 originated from pigs with diarrhea and 2 from pigs without diarrhea. In 14 samples from wild boars, in 8 samples from pigs without diarrhea, and in 3 samples from pigs with diarrhea, neither the mimic nor the target DNA was visible. This indicated the presence of inhibitors in these samples. It is concluded that the mimic can be used as an internal control in the diagnosis of L. intracellularis to indicate inhibition of PCR.

scholarly journals Clinical and epidemiological aspects of canine parvovirus (CPV) enteritis in the State of Rio de Janeiro: 1995 - 2004

2007 ◽  
Vol 59 (2) ◽  
pp. 333-339 ◽  
Author(s):  
T.X. Castro ◽  
S.C. Miranda ◽  
N.V. Labarthe ◽  
L.E. Silva ◽  
R.C.N. Cubel Garcia
Keyword(s):  
Cell Culture ◽  
Polymerase Chain Reaction ◽  
Enzyme Immunoassay ◽  
Hemagglutination Inhibition ◽  
Rio De Janeiro ◽  
Virus Isolation ◽  
Canine Parvovirus ◽  
The State ◽  
Chain Reaction ◽  
Polymerase Chain

This paper relates the clinical and epidemiological aspects of canine parvovirus infection (CPV) in the State of Rio de Janeiro from April 1995 to March 2004. A total of 341 fecal samples were collected from up to 6-months-old puppies with gastroenteritis. The diagnosis of CPV infection was confirmed by hemagglutination/ hemagglutination inhibition tests, enzyme immunoassay, virus isolation in cell culture or polymerase chain reaction. One hundred and fifty-seven samples (46%) were positive for CPV. No correlation among sex, breed or age and the occurrence of CPV infection was observed. The classical signs of parvoviral enteritis (anorexia, lethargy, vomiting and hemorrhagic fluid diarrhea) were observed in 70% of CPV-positive and in 60% of CPV-negative puppies. Although CPV could be detected throughout the studied period, its occurrence was significantly higher from June to September and November to December. These results show that CPV is still circulating in the State of Rio de Janeiro.

scholarly journals Comparison of Three Rapid Commercial Canine Parvovirus Antigen Detection Tests with Electron Microscopy and Polymerase Chain Reaction

2009 ◽  
Vol 21 (3) ◽  
pp. 344-345 ◽  
Author(s):  
Silke Schmitz ◽  
Christina Coenen ◽  
König Matthias ◽  
Thiel Heinz-Jürgen ◽  
Reto Neiger
Keyword(s):  
Electron Microscopy ◽  
Polymerase Chain Reaction ◽  
Chronic Diarrhea ◽  
Gastrointestinal Disease ◽  
Clinical Signs ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Group A ◽  
Polymerase Chain ◽  
Group B

Different antibody-based tests for rapid detection of Canine parvovirus antigens in feces are commercially available, allowing quick diagnosis in a clinical setting. However, the diagnostic accuracy of these tests compared with standard methods has not been evaluated so far. In the current study, 3 commercial tests were compared with immune-electron microscopy (IEM) and polymerase chain reaction (PCR). Dogs were divided into 3 groups: group A, samples from dogs with acute hemorrhagic diarrhea ( n = 50); group B, dogs with chronic diarrhea ( n = 10); and group C, dogs with no evidence of gastrointestinal disease ( n = 40). Specificity of all 3 commercial tests versus PCR and IEM was good to excellent (92.2–100%). Sensitivity, in contrast, was poor: 15.8–26.3% versus PCR and 50–60% versus IEM. In group A, 10 dogs were positive by IEM and 24 dogs were positive by PCR. Positive PCR results were also obtained from animals in control groups (group B, 1 dog; group C, 5 dogs). No dog in group B or C was positive by IEM. In conclusion, the rapid tests are useful to diagnose canine parvoviral enteritis, but they do not rule out parvovirus infection in an animal with typical clinical signs. In addition, a small percentage of healthy dogs and dogs with chronic diarrhea showed positive PCR results; this may be due to asymptomatic/persistent infection or intestinal passage of virus. The significance of this finding remains unclear.

scholarly journals Amplification of human beta-actin gene by the reverse transcriptase-polymerase chain reaction: implications for assessment of RNA from formalin-fixed, paraffin-embedded material.

1996 ◽  
Vol 44 (10) ◽  
pp. 1205-1207 ◽  
Author(s):  
A Dakhama ◽  
V Macek ◽  
J C Hogg ◽  
R G Hegele
Keyword(s):  
Polymerase Chain Reaction ◽  
Pcr Amplification ◽  
Housekeeping Gene ◽  
Chain Reaction ◽  
Viral Genomes ◽  
Negative Results ◽  
Beta Actin ◽  
Target Nucleic Acid ◽  
Formalin Fixed Paraffin Embedded ◽  
Polymerase Chain

The polymerase chain reaction (PCR) is a powerful method that allows enzymatic amplification of rate target nucleic acid sequences. It has been applied to the amplification of viral genomes from paraffin-embedded pathology specimens. However, interpretation of negative results requires amplification of a housekeeping gene such as beta-actin. In the present study we used specific oligonucleotide primers previously designed to amplify both the genomic DNA and the mRNA transcript from paraffin-embedded tissue. These products have predicted sizes of 250 BP and 154 BP, respectively, but our results showed that PCR amplification only (without reverse transcription) unexpectedly generated the 154-BP product. Further investigation of the nature of this product demonstrated that it originated from the amplification of DNA, not RNA. We conclude that the 154-BP product generated by these primers cannot be exclusively considered as beta-actin RNA product and should not be used to assess successful extraction of RNA, to ascertain its integrity, or to normalize for the total amount of RNA assayed by RT-PCR from paraffin-embedded tissue.

Polymerase Chain Reaction Detection of Salmonella spp. in Fecal Samples of Pet Birds

2008 ◽  
Vol 3 (1) ◽  
pp. e29-e29
Author(s):  
B. Sareyyüpoğlu ◽  
A Çelik Ok ◽  
Z. Cantekin ◽  
H. Yardimci ◽  
M. Akan ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Detection ◽  
Chain Reaction ◽  
Salmonella Spp ◽  
Fecal Samples ◽  
Pet Birds ◽  
Polymerase Chain

scholarly journals UTILITY OF ANTIGEN DETECTION TEST AND POLYMERASE CHAIN REACTION IN THE DIFFERENTIATION OF TUBERCULOUS AND NON-TUBERCULOUS MYCOBACTERIA

2017 ◽  
Vol 10 (6) ◽  
pp. 289
Author(s):  
Yogita Singh ◽  
Raji Vasanth ◽  
Shrikala Baliga ◽  
Dhanashree B
Keyword(s):  
Polymerase Chain Reaction ◽  
False Negative ◽  
Diagnostic Methods ◽  
Clinical Samples ◽  
Chain Reaction ◽  
College Hospital ◽  
Medical College ◽  
Negative Results ◽  
Polymerase Chain ◽  
Mycobacterium Spp

Objectives: Cultivation and identification of mycobacteria to species level remains difficult and time-consuming. Hence, easy and rapid diagnostic methods are necessary for the differentiation of Mycobacterium tuberculosis (MTB) from non-tuberculous mycobacteria (NTM). The present study aims to detect and differentiate MTB from NTM isolated from clinical samples by immunochromatographic test (ICT) and polymerase chain reaction (PCR). Methods: Over a period of 1 year, clinical samples (n=496) received from suspected cases of TB, at the Department of Microbiology, Kasturba Medical College Hospital, Mangalore were cultured to isolate Mycobacterium spp. Identification of all the isolates was done by conventional biochemical technique, ICT, and PCR. Results: Among the 496 samples processed, 49 (9.87%) were acid-fast bacilli smear positive and 59 (11.89%) samples showed the growth of Mycobacterium spp. Among these, 10 were rapid growers, 49 were slow-growing mycobacteria, out of which 30 were MTB as identified by conventional biochemical reaction. Out of 59 Mycobacterial isolates subjected to ICT for the detection of MPT 64 antigen, only 28 were identified as MTB. However, all the 30 isolates were correctly identified as MTB by PCR. Conclusion: Hence, PCR is essential for rapid differentiation of non-tuberculous Mycobacterium from MTB. False negative results seen with immunochromatographic MPT 64 antigen assay could be due to mutations within the mpt64 gene. Further studies are necessary to characterize these PCR-positive and immunochromatographic assay negative MTB isolates.

scholarly journals Application of Polymerase Chain Reaction for the Correlation of Salmonella Serovars Recovered from Greyhound Feces with Their Diet

1993 ◽  
Vol 5 (3) ◽  
pp. 378-385 ◽  
Author(s):  
Gregory G. Stone ◽  
M. M. Chengappa ◽  
Richard D. Oberst ◽  
Nathan H. Gabbert ◽  
Scott McVey ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Fecal Material ◽  
Chain Reaction ◽  
Antigenic Analysis ◽  
Fecal Samples ◽  
E Coli ◽  
Staphylococcus Intermedius ◽  
Standard Procedures ◽  
Polymerase Chain ◽  
Salmonella Serovars

The polymerase chain reaction was employed to correlate Salmonella serovars isolated from fecal material of greyhounds suffering from gastroenteritis with those isolated from the diet fed to the greyhounds prior to onset of diarrhea. Kennels around the Abilene, Kansas, area were contacted and supplied with materials needed to collect a portion of the diet each day. With t e onset of diarrhea, the kennels were instructed to ship the fecal material and diet from the previous 10 days to the laboratory for testing. Forty-one fecal samples and corresponding diets were screened for Salmonella, Clostridium perfringens, Campylobacterjejuni, Staphylococcus aureus, Staphylococcus intermedius, and pathogenic (piliated) Escherichia coli by direct culture using standard procedures. The fecal material was also screened for coronavirus and parvovirus using electron microscopy. Thirty-five “normal” fecal samples were screened for all of the above mentioned microorganisms as a control. In addition, the fecal material was screened for E. coli verotoxins I and II and clostridial enterotoxins. A total of 61 Salmonella isolates were recovered from the 41 samples of feces and diet submitted for testing; 31 were recovered from the feces and 30 from the diet. Four Salmonella isolates were recovered from the normal fecal samples. Results obtained by PCR, plasmid profiles, antigenic analysis, and antibiogram profiles indicated that 16 of the 31 isolates recovered from the fecal material were the same strain as that recovered from the diet.

Molecular characterization of canine parvovirus in Brazil by polymerase chain reaction assay

2000 ◽  
Vol 75 (2) ◽  
pp. 127-133 ◽  
Author(s):  
Cesar A.D Pereira ◽  
Telma A Monezi ◽  
Dolores U Mehnert ◽  
Magali D’Angelo ◽  
Edison L Durigon
Keyword(s):  
Polymerase Chain Reaction ◽  
Molecular Characterization ◽  
Polymerase Chain Reaction Assay ◽  
Canine Parvovirus ◽  
Chain Reaction ◽  
Polymerase Chain
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