African Swine Fever (ASF) control is severely limited by a diagnostic gap, as laboratory-based quantitative polymerase chain reaction (qPCR) is highly sensitive but slow, whereas field-deployable immunochromatographic assays (ICAs) are rapid but unreliable. To address this limitation, this study evaluated a novel, rapid isothermal assay, RNase hybridization-assisted amplification (RHAM), as a high-sensitivity, point-of-need diagnostic solution. This study compared the performance of RHAM and a conventional p72-based ICA against the qPCR reference standard using 106 diverse clinical field samples, including oral swabs, blood, serum, and organs, collected from suspected ASF cases in Thailand. The ICA exhibited markedly low diagnostic performance, achieving only 56.76% sensitivity and showing moderate agreement (κ = 0.421) with qPCR, highlighting the need for a more reliable alternative. In contrast, the RHAM assay achieved 94.59% sensitivity and 96.88% specificity, providing results rapidly within 35 min. This statistically superior performance (McNemar’s test, p < 0.0001) demonstrated almost perfect agreement (κ = 0.891) with the qPCR reference standard, missing only four samples with very high Ct values (>30). In conclusion, RHAM is a powerful, accurate, and field-deployable diagnostic tool that effectively bridges the diagnostic gap, offering qPCR-like sensitivity for the rapid containment of ASF outbreaks.

Background and Aim: Serum progesterone concentration plays critical role in determining the optimal breeding time in bitches and diagnosing reproductive-related issues. This study aimed to conduct a comparative analysis of serum progesterone results obtained from commercial point-of-care immunological analyzers, namely, Vcheck®, with those obtained using chemiluminescent microparticle immunoassay (CMIA). Our overarching goal was to evaluate these analyzers’ accuracy and establish standardized guidelines for optimal breeding timing. Materials and Methods: Ninety-four serum samples from bitches were analyzed using the Vcheck® analyzer and compared with CMIA. Thorough documentation included the mean, standard deviation, 95% confidence interval (CI), and minimum and maximum values of serum progesterone concentrations. Furthermore, Pearson’s correlation coefficient, Lin’s concordance correlation coefficient, and the bias correction factor were meticulously recorded. Results: The mean progesterone concentration measured using the Vcheck® analyzer was significantly lower than that measured using CMIA, with a mean difference of 1.26 ng/mL of serum. The Bias correction factor was 0.935, which was nearly 1.00, indicating that the line of best-fit was on the perfect line of agreement, providing insight into the measurement accuracy. Pearson’s correlation coefficient, a measure of precision, was also close to 1 (0.939), confirming the reliability of the data. Furthermore, Lin’s concordance correlation coefficient was 0.877, indicating a fair overall agreement between the Vcheck® and CMIA methods. These results support the validity of the Vcheck® analyzer’s results. The present study was developed by aligning with established CMIA guidelines and adapting them using the range and 95% CI derived from each set of results, ensuring a standardized and rigorous approach. Conclusion: The Vcheck® analyzer provides a rapid assessment of serum progesterone concentration in bitches, with results comparable to those measured using the CMIA technique. However, when considering the use of the Vcheck® analyzer, it is recommended that the results should be interpreted carefully and the interpretation guidelines should be followed. In conclusion, Vcheck® provides a reliable and convenient method for veterinarian practitioners to measure canine progesterone levels in a clinical/hospital setting. Keywords: chemiluminescent microparticle immunoassay, optimal breeding time, progesterone, Vcheck®.

Surgical intervention is essential for many orthopedic conditions in dogs, which are a common cause of pain and debilitation

African Swine Fever (ASF) control is severely hampered by the reliance on slow, laboratory-bound diagnostics. While rapid, field-deployable lateral flow assays (LFAs) are urgently needed, the comparative performance of key single-antigen targets remains poorly characterized. This study aimed to develop and systematically evaluate the diagnostic performance of three in-house single-antigen LFAs targeting ASF virus P30, P54, and P72, using swine field samples from Thailand, including a panel of 143 quantitative polymerase chain reaction-negative swine serum samples. The performance of each LFA was compared against a commercial multi-antigen (P32/P62/P72) indirect ELISA, which served as the reference standard, classifying 64 samples as positive and 79 as negative. The P72-based LFA demonstrated perfect diagnostic performance (100% sensitivity, 100% specificity) and perfect agreement (κ = 1.0) with the enzyme-linked immunosorbent assay (ELISA). Similarly, the P30 LFA demonstrated high performance (100% sensitivity, 98.7% specificity) with ‘Almost Perfect’ agreement (κ = 0.9859). In contrast, the P54 LFA was unsuitable, achieving 100% sensitivity but unacceptably low specificity (88.6%) due to a high rate of false positives. Overall, the single-antigen P72 and P30 LFAs demonstrated excellent concordance with the multi-antigen ELISA, supporting their reliable for detecting antibodies against ASFV. Although these assays do not replace molecular methods for acute infection detection, they represent valuable complementary tools for serosurveillance.

The measurement of serum progesterone often varies due to different laboratory methodologies and individual canine characteristics. In this investigation, serum progesterone outcomes obtained from a commercial point-of-care immunological analyzer, designed for efficient serum progesterone assessment in bitches, were compared with results derived from chemiluminescent microparticle immunoassay from reference laboratories in Thailand. Our thorough documentation encompassed various parameters: mean, standard deviation, 95% confidence interval, and minimum and maximum serum progesterone concentration values. Additionally, we meticulously recorded the Pearson’s correlation coefficient, Lin’s concordance correlation coefficient, and the bias correction factor. Interestingly, there was no significant difference (p > 0.05) in the means obtained by the point-of-care immunological analyzer and chemiluminescent microparticle immunoassay. The Pearson’s correlation coefficient between the point-of-care immunological analyzer and chemiluminescent microparticle immunoassay stood at 0.957, with Lin’s concordance correlation coefficient for point-of-care immunological analyzer recorded as 0.949. Furthermore, the bias correction factor was established at 0.991. This investigation followed established chemiluminescent microparticle immunoassay guidelines, modified to incorporate the mean and 95% confidence interval as criteria for optimal breeding time using the point-of-care immunological analyzer. In conclusion, the commercial point-of-care immunological analyzer emerges as a valuable tool, aiding in precisely determining the optimal timing for natural mating or artificial insemination in bitches.

The measurement of sP4 often varies due to different laboratory methodologies and individual canine characteristics. In this investigation, sP4 outcomes obtained from a commercial POC analyzer, designed for efficient sP4 assessment in bitches, were compared with results derived from CMIA, the reference laboratories in Thailand. Thorough documentation encompassed various parameters: mean, standard deviation (SD), 95% confidence interval (CI), minimum and maximum sP4 concentration values. Additionally, we meticulously recorded the Pearson’s correlation coefficient, Lin’s concordance correlation coefficient, and the bias correction factor. Interestingly, there was no significant difference (p > 0.05) in the means obtained by the POC and CMIA. The Pearson’s correlation coefficient between POC and CMIA stood at 0.957, with Lin’s concordance correlation coefficient for POC recorded as 0.949. Furthermore, the bias correction factor was established at 0.991. This investigation followed established CMIA guidelines, modified to incorporate the mean and 95% CI as criteria for optimal breeding time using the POC analyzer. In conclusion, the commercial POC immunologic analyzer emerges as a valuable tool, aiding in precisely determining the optimal timing for natural mating or artificial insemination in bitches.