Evaluation of polymerase chain reaction and dot blot hybridisation tests in the diagnosis of pigeon circovirus infections

Todd, D.; Duchatel, Jean-Pierre; Weston, Jonathan; Ball, N W; Borghmans, B. J.; Moffett, D.; Smyth, Joan A.

doi:10.1016/s0378-1135(02)00154-2

Cited by 42 publications

(41 citation statements)

References 11 publications

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“…When amplification was performed in the presence of negative duck BF cellular DNA, conditions that more closely resemble those occurring in extracts of clinical samples, the limit of detection of the conventional test was reduced by 100-fold and that of the real-time test was reduced 10-fold, indicating that the real-time test was potentially more sensitive. Nevertheless, the conventional PCR test was considered to have greater sensitivity than tests we developed previously to detect PiCV (Todd et al ., 2002) and GoCV (Ball et al ., 2004). The specificity of the conventional PCR test was confirmed when pigeon and goose BF extracts, which were positive using PiCV and GoCV PCR tests, respectively, were found to be negative by the conventional DuCV PCR test.…”

Section: Discussionmentioning

confidence: 99%

“…The PCR products were separated by electrophoresis on a 2% (w/v) of agarose gel and the DNA bands were stained with ethidium bromide (0.5 mg/ml) and visualized using ultraviolet light. The specificity of the test was evaluated by applying the PCR test to known GoCV-positive and PiCV-positive samples of BF (Todd et al ., 2002;Ball et al ., 2004), respectively. SYBR Green real-time PCR test.…”

Section: Methodsmentioning

confidence: 99%

“…This paper describes the first development of a PCR test for diagnosing DuCV infections and its use in assessing the prevalence of circovirus infections in dead and sick ducks. In the absence of serological tests, PCR has proved to be a specific and sensitive method for diagnosing avian circovirus infections (Ypelaar et al ., 1999;Todd et al ., 2002;Ball et al ., 2004). However, its ability to detect circovirus DNA in clinically normal birds, for example pigeons (Todd et al ., 2002), suggests that while it may be useful for diagnosing circovirus infections, PCR may be too sensitive to use as a diagnostic of disease.…”

Section: Introductionmentioning

confidence: 99%

“…In the absence of serological tests, PCR has proved to be a specific and sensitive method for diagnosing avian circovirus infections (Ypelaar et al ., 1999;Todd et al ., 2002;Ball et al ., 2004). However, its ability to detect circovirus DNA in clinically normal birds, for example pigeons (Todd et al ., 2002), suggests that while it may be useful for diagnosing circovirus infections, PCR may be too sensitive to use as a diagnostic of disease. This has prompted speculation that, in terms of clinical severity, the effects of circovirus infection may depend on virus load (Todd et al ., 2002;Ball et al ., 2004).…”

Section: Introductionmentioning

confidence: 99%

“…However, its ability to detect circovirus DNA in clinically normal birds, for example pigeons (Todd et al ., 2002), suggests that while it may be useful for diagnosing circovirus infections, PCR may be too sensitive to use as a diagnostic of disease. This has prompted speculation that, in terms of clinical severity, the effects of circovirus infection may depend on virus load (Todd et al ., 2002;Ball et al ., 2004). With the aim of investigating the circovirus loads in infected ducks, this present paper also reports the development of a real-time PCR test, based on SYBR Green chemistry, and its application to selected BF samples.…”

Section: Introductionmentioning

confidence: 99%

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Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests

et al. 2005

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Development of the first conventional and real-time polymerase chain reaction (PCR) tests for the diagnoses of duck circovirus (DuCV) infections is described. Both tests amplified a 230 bp fragment specific to the DuCV Rep gene. Although both tests had the same detection limit (13 )/10 3 target DNA/ml) when the target DNA was diluted in water, the detection limit of the real-time test (13 )/10 4 target DNA/ml) was 10-fold less than the conventional test (13)/10 5 target DNA/ml) when the amplifications were performed in the presence of cellular DNA. Using the conventional PCR test, DuCV DNA was detected in 85 (84%) of 101 bursa of Fabricius samples from dead or sick ducks, aged between 1 and 12 weeks, and in samples from 35 (94%) of 37 flocks. Application of the SYBR Green-based real-time PCR test to 54 selected bursa of Fabricius samples indicated that more samples were positive by real-time PCR than by conventional PCR, allowed the numbers of genome copies to be estimated and showed that some bursa of Fabricius samples contained over 10 13 genome copies/g tissue. Although DuCV infections were detected in birds aged from 1 to 12 weeks, higher virus DNA levels were detected in ducks aged older than 5 weeks than in ducks younger than 5 weeks. An in situ hybridization method for the detection of DuCV in histological samples was also developed. Additional work is required to determine the clinicopathological significance of DuCV infections.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests

et al. 2005

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Chicken Infectious Anemia Virus and Other Circovirus Infections

Schat

Woods

2013

Diseases of Poultry

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Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus

et al. 2013

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There are no effective antiviral treatments for pigeon circovirus (PiCV); thus, rapid diagnosis is critical for effective control of the disease caused by this virus. The recent development of a novel LAMP technique that amplifies nucleic acids rapidly with high specificity and sensitivity under isothermal conditions has overcome some of the deficiencies of nucleic-acid-based diagnostic tests. We established a LAMP method for rapid detection of PiCV using two pairs of primers that were designed from PiCV and compared its sensitivity and specificity with that of PCR. Amplification by LAMP was optimal at 63 °C for 60 min. The detection limit was nearly 0.5 pg of PiCV DNA, making it ten times more sensitive than PCR. There was no cross-reaction with porcine circovirus type 2 (PCV2), pigeon Trichomonas gallinae, or pigeon herpesvirus (PHV) under the same conditions. The assay also successfully detected the pathogen DNA in the tissues of infected pigeons. This is the first report indicating that LAMP is a valuable, rapid method of detecting PiCV with high sensitivity and specificity.

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Evaluation of polymerase chain reaction and dot blot hybridisation tests in the diagnosis of pigeon circovirus infections

Cited by 42 publications

References 11 publications

Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests

Diagnosis of duck circovirus infections by conventional and real-time polymerase chain reaction tests

Chicken Infectious Anemia Virus and Other Circovirus Infections

Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus

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