TH22252U - Primers for detecting the presence of APP in pigs. and methods for detecting infection using primers - Google Patents

Abstract

17/07/2020(OCR) This invention is the development of a primer for detecting infection. Actinobacillus pleuropneumoniae or APP virus To diagnose lung disease and pleurisy in pigs. The primers are highly specific to the genetic material of the APP strain. It is also highly sensitive even when the temperature at which the primers bind to the initial DNA strand increases. As a result, the process can be reduced and the time taken for diagnosing lung disease and pleurisy in pigs using the polymerase chain reaction method without compromising the specificity or sensitivity of the reaction. Page 1 of number 1 page summary of invention This invention is the development of a primer for detecting infection. Actinobacillus pleuropneumoniae or APP bacteria for diagnosis of lung disease and pleurisy in pigs. Such primers are highly specific to Genetic material of APP germs It is also highly sensitive even when the temperature at which the primers can bind to the initial DNA strand increases. As a result, the process can be shortened and the time taken for diagnosing lung disease and pleurisy will be reduced. in pigs by polymerase chain reaction without losing the specificity or sensitivity of the reaction.

Claims (6)

1. /07/2020(OCR) 1. Primers for detecting APP in pigs consist of at least one primer from The first primer (P25) having the nucleotide sequence 5' ATGTCACCGGCAAAGGGAATGATGTGC 3', the second primer (P26) having the nucleotide sequence 5' CTCTACCCGCTTAGGATCCGCC 3', the third primer (P27) having the nucleotide sequence 5' GAGGATTTGTTTCTCGGTGGTGAGGG 3', the fourth primer (P28) having the nucleotide sequence 5' TCGCCGTTTTCTCTTCCCAA 3', the fifth primer (P29) having the nucleotide sequence 5' TGCGCCAAATAACCCGGGACGTAAC 3', or the sixth primer (P30) having the nucleotide sequence 5' TCATCCGCAGCGAGCCAGCC 3'. 2. The primer according to claim 1, wherein said primers 3. The primer of claim 1, wherein said primer is used in a PCR technique. 4. The primer of claim 1, wherein said primer is capable of binding to the starting DNA strand by a PCR technique at a temperature in the range of 5575°C. 5. A method for detecting APP in a biological specimen of a pig using the primers of claim 1, having the steps as follows: a. taking the DNA extracted from the biological specimen of the pig and subjecting it to a PCR technique by selecting at least one primer from The first primer (P25) with the nucleotide sequence 5' ATGTCACCGGCAAAGGGAATGATGTGC 3', the second primer (P26) with the nucleotide sequence 5' CTCTACCCGCTTAGGATCCGCC 3', the third primer (P27) with the nucleotide sequence 5' GAGGATTTGTTTCTCGGTGGTGAGGG 3', the fourth primer (P28) with the nucleotide sequence 5' TCGCCGTTTTCTCTTCCCAA 3', the fifth primer (P29) with the nucleotide sequence 5' TGCGCCAAATAACCCGGGACGTAAC 3', or the sixth primer (P30) with the nucleotide sequence 5' TCATCCGCAGCGAGCCAGCC 3', by PCR performed under conditions with a temperature at which the primers can bind to the starting DNA strand within the range of 5575 °C. To obtain a PCR product, b. Analysis of the PCR product by electrophoresis. 6. The method of claim 5, wherein the biological sample of the pig is selected from any or a combination of tissues, blood, serum, secretions, etc. Page 1 of 1 Claim 1. The primers for detecting APP in pigs comprise at least one primer from Primer one (P25) with the nucleotide sequence 5' ATGTCACCGGCAAAGGGAATGATGTGC 3', primer two (P26) with the nucleotide sequence 5' CTCTACCCGCTTAGGATCCGCC 3', primer three (P27) with the nucleotide sequence 5' GAGGATTTGTTTCTCGGTGGTGAGGG 3', primer four (P28) with the nucleotide sequence 5' TCGCCGTTTTCTCTTCCCAA 3', primer five (P29) with the nucleotide sequence 5' TGCGCCAAATAACCCGGGACGTAAC 3', or primer six (P30) with the nucleotide sequence 5' TCATCCGCAGCGAGCCAGCC 3'. 2. A primer according to claim 1, wherein said primer is specific to the apxIVA gene. 3. A primer according to claim 1, wherein said primer is used in the PCR technique. 4. The primer according to claim 1, wherein said primer can bind to the starting DNA strand by PCR technique at a temperature in the range of 5575°C. 5. A method for detecting APP in a biological sample of pigs using primers according to claim 1, wherein the steps are as follows: (a) taking the DNA extracted from the biological sample of pigs and performing a PCR technique by selecting at least one primer from The first primer (P25) with the nucleotide sequence 5' ATGTCACCGGCAAAGGGAATGATGTGC 3', the second primer (P26) with the nucleotide sequence 5' CTCTACCCGCTTAGGATCCGCC 3', the third primer (P27) with the nucleotide sequence 5' GAGGATTTGTTTCTCGGTGGTGAGGG 3', the fourth primer (P28) with the nucleotide sequence 5' TCGCCGTTTTCTCTTCCCAA 3', the fifth primer (P29) with the nucleotide sequence 5' TGCGCCAAATAACCCGGGACGTAAC 3', or the sixth primer (P30) with the nucleotide sequence 5' TCATCCGCAGCGAGCCAGCC 3', by PCR performed under conditions with a temperature at which the primers can bind to the starting DNA strand within the range of 5575 °C. To obtain PCR products, b. Analysis of PCR products by electrophoresis. 6. The method of claim 5, wherein the biological sample of a pig is selected from any or a combination of tissue, blood, serum, secretions, or other material.