NDLI: Establishment of a Direct PCR Assay for Simultaneous Differential Diagnosis of African Swine Fever and Classical Swine Fever Using Crude Tissue Samples

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Establishment of a Direct PCR Assay for Simultaneous Differential Diagnosis of African Swine Fever and Classical Swine Fever Using Crude Tissue Samples

Content Provider	MDPI
Author	Nishi, Tatsuya Okadera, Kota Fukai, Katsuhiko Yoshizaki, Miwa Nakasuji, Ai Yoneyama, Syuji Kokuho, Takehiro
Copyright Year	2022
Description	African swine fever (ASF) and classical swine fever (CSF) are contagious swine diseases that are clinically indistinguishable from each other; hence, reliable test methods for accurate diagnosis and differentiation are highly demanded. By employing a buffer system suitable for crude extraction of nucleic acids together with an impurity-tolerant enzyme, we established a multiplex assay of real-time reverse-transcription polymerase chain reaction (rRT-PCR) for simultaneous detection of ASF virus (ASFV), CSF virus (CSFV) and swine internal control derived genes in a sample without the need for prior purification of viral nucleic acids. We applied this method to test serum and tissue samples of infected pigs and wild boars and compared the statistical sensitivities and specificities with those of standard molecular diagnostic methods. When a serum was used as a test material, the newly established assay showed 94.4% sensitivity for both and 97.9 and 91.9% specificity for ASFV and CSFV detection, respectively. In contrast, the results were 100% identical with those obtained by the standard methods when a crude tissue homogenate was used as a test material. The present data indicate that this new assay offers a practical, quick, and reliable technique for differential diagnosis of ASF and CSF where geographical occurrences are increasingly overlapping.
Starting Page	498
e-ISSN	19994915
DOI	10.3390/v14030498
Journal	Viruses
Issue Number	3
Volume Number	14
Language	English
Publisher	MDPI
Publisher Date	2022-02-28
Access Restriction	Open
Subject Keyword	Viruses Infectious Diseases Microbiology African Swine Fever Classical Swine Fever Real-time Reverse-transcription Polymerase Chain Reaction Multiplex Detection Crude Nucleic Acid Extraction
Content Type	Text
Resource Type	Article

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