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J Vet Med Sci ; 84(9): 1157-1163, 2022 Sep 01.
Article in English | MEDLINE | ID: covidwho-2021433


Infectious bronchitis virus (IBV) is the causative agent of infectious bronchitis (IB) in chickens. There is a correlation between cross-protection and percentage of similarity between nucleotide sequences encoding the S1 subunit, which is responsible for generating neutralizing and serotype-specific antibodies. Therefore, RT-PCR is commonly used to amplify the IBV-S1 gene following DNA sequencing in order to predict the efficacy of vaccines against IBV strains. We successfully enhanced the sensitivity for detection of the IBV-S1 gene by second PCR after purification of the 1st RT-PCR product. Using that method, we obtained detailed information on the prevalence of IBV on poultry farms in Gifu Prefecture, Japan. The IBV-S1 gene detection method used in the current study will enable accurate information on the prevalence of IBV in Japan to be obtained.

Coronavirus Infections , Infectious bronchitis virus , Poultry Diseases , Animals , Chickens , Coronavirus Infections/diagnosis , Coronavirus Infections/epidemiology , Coronavirus Infections/veterinary , Farms , Japan/epidemiology , Poultry , Poultry Diseases/diagnosis , Poultry Diseases/epidemiology , Poultry Diseases/prevention & control , Reverse Transcriptase Polymerase Chain Reaction/veterinary
Avian Dis ; 64(2): 149-156, 2020 06.
Article in English | MEDLINE | ID: covidwho-892406


Infection of the oviduct by an infectious bronchitis virus (IBV) in laying hens has been associated with the false layer syndrome. Because the diagnostic procedure for the detection of cystic oviducts by postmortem examinations in IBV-positive replacement pullet flocks could involve the unnecessary sacrifice of numerous healthy pullets without reproductive tract anomalies, the development of a noninvasive and nonlethal diagnostic procedure would be desirable. The first objective of the study was to evaluate the diagnostic accuracy of a transcutaneous ultrasonography method to predict the presence of cystic oviducts compared to postmortem examinations in a commercial pullet flock positive for an IBV genotype Delmarva (DMV) variant. The second objective was to evaluate the performance of the same ultrasonography method to later detect false layers in the same flock in sexually mature hens by identifying the presence of an egg in the oviduct due to the presence of atretic oviducts undetectable by ultrasonography and the absence of cystic oviducts at that age. In replacement pullets, the sensitivity (Se) and specificity (Sp) of the ultrasonography (index test) compared to the postmortem examination (reference standard test) were 73% and 91%, respectively. The positive predictive value (PPV) and negative predictive value (NPV) were 67% and 93%. The ultrasonography technique showed a positive likelihood ratio (LR+) of 7.82 and a negative likelihood ratio (LR-) of 0.30. In sexually mature hens, the Se, Sp, PPV, and NPV of the ultrasonography compared to the laying status were 98%. The LR+ was 49.00 and the LR- was 0.02 when compared to the laying status. In conclusion, the ultrasonography could replace postmortem examinations to detect cystic oviducts in commercial flocks of replacement pullets previously infected with an IBV-DMV 1639 variant. Although the test accuracy of ultrasonography was excellent for the hens at production peak to identify laying and nonlaying hens based on the presence of an egg in the reproductive tract, its practicality was limited due to atretic oviducts being not detectable.

Precisión diagnóstica de la ultrasonografía para detectar gallinas falsas ponedoras en una parvada comercial infectada por un virus de la bronquitis infecciosa genotipo Delmarva que causa oviductos quísticos. La infección del oviducto por el virus de bronquitis infecciosa (IBV) en gallinas de postura se ha asociado con el síndrome de la falsa ponedora. Debido a que el procedimiento de diagnóstico para la detección de oviductos quísticos mediante exámenes post mortem en parvadas de pollitas de reemplazo positivas para bronquitis infecciosa podría involucrar el sacrificio innecesario de numerosas pollitas sanas sin anomalías del tracto reproductivo, por lo tanto es deseable el desarrollo de un procedimiento de diagnóstico no invasivo y no letal. El primer objetivo del estudio fue evaluar la precisión diagnóstica de un método de ultrasonografía transcutánea para predecir la presencia de oviductos quísticos en comparación con los exámenes post mortem en un lote comercial de pollitas que resultó positivo para una variante del genotipo Delmarva (DMV) del virus de la bronquitos infecciosa. El segundo objetivo fue evaluar el desempeño del mismo método de ultrasonografía para detectar posteriormente gallinas falsas en la misma parvada en las gallinas sexualmente maduras mediante la identificación de la presencia de un huevo en el oviducto debido a la presencia de oviductos atrésicos indetectables por ultrasonografía y la ausencia de oviductos quísticos a esa edad. En las pollitas de reemplazo, la sensibilidad (Se) y la especificidad (Sp) de la ultrasonografía (prueba de índice) en comparación con el examen post mortem (prueba estándar de referencia) fueron de 73% y 91%, respectivamente. El valor predictivo positivo (VPP) y el valor predictivo negativo (VPN) fueron 67% y 93%. La técnica de ultrasonografía mostró una razón de probabilidad positiva (LR+) de 7.82 y una razón de probabilidad negativa (LR­) de 0.30. En las gallinas sexualmente maduras, la Se, Sp, PPV y NPV de la ultrasonografía en comparación con el estado de postura fueron del 98%. El LR + fue 49.00 y el LR­fue 0.02 en comparación con el estado de la postura. En conclusión, la ultrasonografía podría reemplazar los exámenes post mortem para detectar oviductos quísticos en parvadas comerciales de pollitas de reemplazo previamente infectadas con una variante DMV-1639 del virus de la bronquitis infecciosa. Aunque la precisión de la prueba de la ecografía fue excelente para las gallinas en el pico de producción para identificar gallinas ponedoras y no ponedoras en función de la presencia de un huevo en el tracto reproductivo, su funcionalidad fue limitada debido a que los oviductos atrésicos no fueron detectables.

Chickens , Coronavirus Infections/veterinary , Cysts/veterinary , Infectious bronchitis virus/isolation & purification , Poultry Diseases/diagnosis , Ultrasonography/veterinary , Animals , Coronavirus Infections/diagnosis , Coronavirus Infections/virology , Cysts/virology , Female , Oviducts/virology , Poultry Diseases/virology , Ultrasonography/statistics & numerical data
Poult Sci ; 99(10): 4809-4813, 2020 Oct.
Article in English | MEDLINE | ID: covidwho-864272


In this study, specific primers and fluorescent probes were designed to target the thymidine kinase (TK) gene sequence of avian infectious laryngotracheitis virus (ILTV). Through specificity and sensitivity tests, a real-time fluorescence-based recombinase-aided amplification (RF-RAA) method for detecting ILTV was established. The results showed that the method was specific and could be used to accurately detect ILTV, and there was no cross-reaction with Newcastle disease virus (NDV), avian influenza virus (AIV), or infectious bronchitis virus (IBV). Real-time fluorescence-based recombinase-aided amplification had high sensitivity, and the lowest detectable limit (LDL) for ILTV could reach 10 copies/µL, 1,000 times more sensitive than conventional PCR (104 copies/µL), to rival that of real-time fluorescence-based quantitative PCR (RFQ-PCR) (10 copies/µL). This method and RFQ-PCR were used to detect 96 samples of chicken throat swabs with ILT initially diagnosed in clinic from the north of China, and the coincidence rate of the 2 methods was 100%. The RF-RAA reaction required only 20-30 minutes to completing, and its sensitivity was much higher than that of conventional PCR. Real-time fluorescence-based recombinase-aided amplification is similar to RFQ-PCR and has the advantages of specificity, sensitivity, and high efficiency, so it is suitable for early clinical detection and epidemiological investigation of ILTV.

Herpesviridae Infections , Herpesvirus 1, Gallid , Poultry Diseases , Animals , Chickens , China , Fluorescence , Herpesviridae Infections/diagnosis , Herpesviridae Infections/veterinary , Herpesviridae Infections/virology , Herpesvirus 1, Gallid/genetics , Limit of Detection , Nucleic Acid Amplification Techniques/veterinary , Poultry Diseases/diagnosis , Poultry Diseases/virology , Recombinases/metabolism , Sensitivity and Specificity