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CN-121992120-A - Primer pair, kit and method for detecting chlamydia pneumoniae

CN121992120ACN 121992120 ACN121992120 ACN 121992120ACN-121992120-A

Abstract

The invention provides a primer pair, a primer set and a primer pair method for detecting chlamydia pneumoniae, wherein the primer pair comprises a forward primer and a reverse primer aiming at a peptidoglycan-related lipoprotein (Pal) gene, and the Pal gene comprises a nucleotide sequence shown as SEQ ID NO. 15. The forward primer is 15 to 25 nucleotides in length and corresponds to the nucleotide sequence between positions 1 and 33 shown in SEQ ID NO. 15. The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 of SEQ ID NO. 15. The invention also provides a kit for detecting the pneumonectasis, comprising the primer pair, and a method for detecting the pneumonectasis by using the primer pair.

Inventors

  • CHEN WEIFAN
  • SHEN YUHAN
  • ZHANG XIUHUI

Assignees

  • 台达电子工业股份有限公司

Dates

Publication Date
20260508
Application Date
20241108

Claims (19)

  1. 1. A primer pair for detecting chlamydia pneumoniae, comprising a forward primer and a reverse primer for a peptidoglycan-associated lipoprotein (Peptidoglycan-Associated Lipoprotein, pal) gene, wherein the Pal gene comprises a nucleotide sequence shown as SEQ ID No. 15; Wherein the forward primer has a length of 15 to 25 nucleotides and corresponds to the nucleotide sequence between the 1 st and 33 th positions shown in SEQ ID NO.15, and The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 of SEQ ID NO. 15.
  2. 2. The primer pair for detecting chlamydia pneumoniae according to claim 1, wherein the forward primer is 19 to 25 nucleotides in length and/or the reverse primer is 18 to 27 nucleotides in length.
  3. 3. The primer pair for detecting chlamydia pneumoniae according to claim 1, wherein the forward primer comprises a nucleotide sequence as set forth in any one of SEQ ID NOs 1-3.
  4. 4. The primer pair for detecting chlamydia pneumoniae according to claim 1, wherein the reverse primer comprises a nucleotide sequence as shown in any one of SEQ ID NOs 4 to 10.
  5. 5. A kit for detecting a chlamydia pneumoniae, comprising: A forward primer, a reverse primer and a probe, wherein the forward primer, the reverse primer and the probe are directed against peptidoglycan-associated lipoprotein (Pal) genes, and the Pal genes comprise a nucleotide sequence shown as SEQ ID NO. 15; Wherein the forward primer has a length of 15 to 25 nucleotides and corresponds to the nucleotide sequence between the 1 st and 33 th positions shown in SEQ ID NO.15, and The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 shown in SEQ ID NO. 15.
  6. 6. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the forward primer is 19 to 25 nucleotides in length and/or the reverse primer is 18 to 27 nucleotides in length.
  7. 7. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the forward primer comprises a nucleotide sequence as set forth in any one of SEQ ID NOs 1 to 3.
  8. 8. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the reverse primer comprises a nucleotide sequence as set forth in any one of SEQ ID NOs:4 to 10.
  9. 9. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the probe has a length of 20 to 30 nucleotides.
  10. 10. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the probe comprises a nucleotide sequence as shown in SEQ ID NO. 11.
  11. 11. The kit for detecting a chlamydia pneumoniae according to claim 5, wherein the 5 'end of the probe is linked to a reporter dye and the 3' end of the probe is linked to a quencher.
  12. 12. The kit for detecting a pneumophila of claim 5, further comprising a forward primer and a reverse primer for a nucleic acid sequence of a human gene, wherein the forward primer comprises a nucleotide sequence shown as SEQ ID NO. 12 and the reverse primer comprises a nucleotide sequence shown as SEQ ID NO. 13.
  13. 13. A method of detecting a chlamydia pneumoniae comprising the steps of: Providing a sample; Providing a primer pair comprising a forward primer and a reverse primer for a peptidoglycan-associated lipoprotein (Pal) gene, wherein the Pal gene comprises the nucleotide sequence shown in SEQ ID No. 15; Wherein the forward primer has a length of 15 to 25 nucleotides and corresponds to the nucleotide sequence between the 1 st and 33 th positions shown in SEQ ID NO.15, and The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 shown in SEQ ID NO. 15; Performing a polymerase chain reaction with the sample using the primer pair to obtain a product, and The product was analyzed to detect the presence of c.
  14. 14. The method of claim 13, further comprising providing a probe, wherein the probe is 20 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 34 and 63 as set forth in SEQ ID No. 15.
  15. 15. The method for detecting a c.pneumoniae according to claim 14 wherein the probe comprises a nucleotide sequence as shown in SEQ ID No. 11.
  16. 16. The method of claim 14, wherein the 5 'end of the probe is linked to a reporter and the 3' end of the probe is linked to a quencher.
  17. 17. The method of detecting a pneumophila of claim 13, wherein the source of the sample comprises a saliva sample, a sputum sample, a nasal swab (nose swab) sample, a throat swab (throat swab) sample, a nasopharynx (nasopharyngeal) sample, a urine sample, a fecal sample, a rectal swab (rectal swab) sample, a cerebrospinal fluid (cerebrospinal fluid, CSF) sample, or a body fluid (body fluid) sample.
  18. 18. The method of claim 13, wherein the step of performing the polymerase chain reaction with the sample using the primer pair to obtain the product comprises performing the polymerase chain reaction such that the primer pair amplifies a nucleotide sequence of the Pal gene in the chlamydia pneumoniae to obtain the product.
  19. 19. The method of claim 13, further comprising providing a forward primer and a reverse primer for a nucleic acid sequence of a human gene, wherein the forward primer comprises a nucleotide sequence as set forth in SEQ ID No. 12 and the reverse primer comprises a nucleotide sequence as set forth in SEQ ID No. 13.

Description

Primer pair, kit and method for detecting chlamydia pneumoniae Technical Field The present invention relates to primer pairs, kits and methods for detecting Chlamydia spp, and more particularly to primer pairs, kits and methods for detecting Chlamydia pneumoniae CHLAMYDIA PNEUMONIAE. Background The pneumophila (CHLAMYDIA PNEUMONIAE) is a common respiratory disease source and is also the main cause of atypical pneumonia infection, and has no specific epidemic season, can be infected all the year round, and has one pandemic every 10 years. Infection with the bacterium Chlamydia pneumoniae is spread worldwide and accounts for about 20% of the cell-infected pneumonia. From serological statistics, more than about half of the population has the presence of antibodies to c. Perennial symptoms caused by the bacteria of the genus Chlamydia are pharyngolaryngitis, others include rhinitis, hoarseness, fever, abdominal pain, diarrhea, nausea, vomiting, etc. Recent studies have also shown that asthma, arteriosclerotic diseases and multiple sclerosis are highly correlated with infection by c. The infection of the bacteria on the garment of the pneumonia is a main pathogen causing the respiratory tract infection of infants and asthma among the infant groups. Atypical pneumonia turns on pathogenic mechanisms, is liable to affect whole body organs, causes multiple organ dysfunction, and causes drop of white blood cell number and liver dysfunction. Typically, patients are most refractory to antibiotic therapy within five days. Since the symptoms of atypical pneumonia are highly similar to influenza or common cold, they are not easily diagnosed early, and thus the opportunity for early treatment is often missed. Therefore, the identification and detection of the chlamydia pneumoniae has important significance for early diagnosis, early treatment and infection risk control of atypical pneumonia. In view of the above, development of a detection tool and a method capable of rapidly and accurately detecting whether the infection is caused by the pathogen of the chlamydia pneumoniae is important for improving the cure rate of the disease, which is one of the problems of the current research in the industry. Disclosure of Invention According to some embodiments of the present invention, there is provided a primer pair for detecting chlamydia pneumoniae, comprising a forward primer and a reverse primer for a peptidoglycan-associated lipoprotein (Pal) gene, and the Pal gene comprises a nucleotide sequence as shown in SEQ id No. 15. The forward primer is 15 to 25 nucleotides in length and corresponds to the nucleotide sequence between positions 1 and 33 shown in SEQ ID NO. 15. The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 of SEQ ID NO. 15. According to some embodiments of the present invention, there is also provided a kit for detecting chlamydia pneumoniae, comprising a forward primer, a reverse primer and a probe. The forward primer, the reverse primer and the probe are directed against peptidoglycan-associated lipoprotein (Pal) genes, and the Pal genes comprise a nucleotide sequence shown as SEQ ID NO. 15. The forward primer is 15 to 25 nucleotides in length and corresponds to the nucleotide sequence between positions 1 and 33 shown in SEQ ID NO. 15. The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 shown in SEQ ID NO. 15. According to some embodiments of the present invention, there is also provided a method of detecting chlamydia pneumoniae comprising providing a sample and providing a primer pair comprising a forward primer and a reverse primer for a peptidoglycan-associated lipoprotein (Pal) gene, and the Pal gene comprises a nucleotide sequence as shown in SEQ ID No. 15. The forward primer is 15 to 25 nucleotides in length and corresponds to the nucleotide sequence between positions 1 and 33 shown in SEQ ID NO. 15. The reverse primer is 15 to 30 nucleotides in length and corresponds to the nucleotide sequence between positions 64 and 123 shown in SEQ ID NO. 15. The method of detecting a pneumophila further comprises performing a polymerase chain reaction with the sample using the primer pair to obtain a product, and analyzing the product to detect the presence of the pneumophila. In order to make the features and advantages of the present invention more comprehensible, several embodiments accompanied with figures are described in detail below. Drawings FIG. 1 shows a partial fragment of the peptidoglycan-associated lipoprotein (Pal) gene sequence (GenBank: LN 847257.1) and the design positions of forward, probe, and reverse primers according to some embodiments of the invention; FIG. 2 shows a temperature profile of a real-time quantitative polymerase chain reaction in accordance with some embodiments of the present invention; FIG. 3A shows an amplification curve of a real-ti