Detection and Segregation of Brucella abortus and Brucella melitensis in Aborted Bovine, Ovine, Caprine, Buffaloes and Camelid Fetuses by Application of Conventional and Real-time Polymerase Chain Reaction

บทคดยอ การศกษานใชปฏกรยาพซอาร แบบ conventional และ real time ในการตรวจสอบและคดแยกเชอ Brucella abortus และ Brucella melitensis ในตวออนแทงของโค แกะ แพะ กระบอ และอฐ ตวอยางทงหมดถกเกบและขนสงไปยงหองปฏบตการทนท เพอทำ การสกดจโนมกดเอนเอ และตรวจดวยปฏกรยาพซอาร แบบ conventional และ real time ดวยไพรเมอรทเฉพาะเจาะจงสำหรบ Brucella abortus และ Brucella melitensis ตวอยางดเอนเอจำนวน 3710 ตวอยางทเกบจากของเหลวในกระเพาะแทของตวออนทแทง ทำการ วเคราะหดวย TaqMan และปฏกรยาพซอาร แบบขนเดยว พบวาจำนวนตวอยางทใหผลบวกตอ Brucella จากตวอยางทงหมดเปน 281/892 (31.5%) ในโค 224/810 (27.65%) ในแกะ 219/786 (27.86%) ในแพะ 199/604 (32.94%) ในกระบอ และ 201/618 (32.52%) ในอฐ เมอใชปฏกรยาพซอาร แบบ conventional และพบวาตวอยางทใหผลบวกตอ B. melitensis และ B. abortus จาก ตวอยางทงหมดคดเปน 45/281 และ 231/281 ในโค 169/224 และ 49/224 ในแกะ 194/219 และ 22/219 ในแพะ 57/199 และ 137/199 ในกระบอ และ 51/201 และ 143/201 ในอฐ เมอใชปฏกรยาพซอาร แบบ real time ความไวและความจำเพาะของ ปฏกรยาพซ อาร แบบ real time คดเปน 100% และ 100% การวเคราะหทางสถตแสดงใหเหนความแตกตางอยางมนยสำคญ (p<0.01) ระหวาง B. abortus และ B. melitensis ทตรวจพบในตวอยางทเกบจากของเหลวในกระเพาะแทของตวออนทแทงของโค แกะ แพะ กระบอ และอฐ บฟฟาโลและทารก Camelid และระหวาง Brucella spp. ในโค กบแพะ กระบอ และอฐ (p<0.05) คา CT ทไดจากปฏกรยาพซอาร แบบ real-time มความแตกตางทางสถตระหวางโค แกะ แพะ กรบอ และอฐสำหรบการตรวจพบ B. abortus และ B. melitensis ผลการศกษา ชใหเหนวาปฏกรยาพซอาร แบบ real-time ใหผลการตรวจเรวกวาวธมาตรฐานในปจจบน ในการแยกและการคดแยกเชอ Brucella spp. คำสำคญ : ตวออนแทง Brucella abortus Brucella melitensis ปฏกรยาพซอาร แบบ conventional ปฏกรยาพซอาร แบบ real-time Abstract This present study was carried out to use conventional and real-time PCR for detection and segregation of Brucella abortus and Brucella melitensis in aborted bovine, ovine, caprine, buffalo and camel fetuses. All samples were collected and immediately transferred to laboratory, genomic DNA was extracted and the conventional and real-time PCR by specific primers for Brucella abortus and Brucella melitensis was performed. A TaqMan analysis and single-step PCR was carried out in total 3710 DNA of abomasal contents of aborted fetuses. In total, 281/892 (31.5%) bovine, 224/810 (27.65%) ovine, 219/786 (27.86%) caprine, 199/604 (32.94%) buffalo and 201/618 (32.52%) camel fetus samples gave positive results for Brucella species by conventional PCR. Moreover, 45/281 and 231/281, 169/224 and 49/224, 194/219 and 22/219, 57/199 and 137/199 and finally 51/201 and 143/201 specimens were positive for B. melitensis and B. abortus in aborted bovine, ovine, caprine, buffalo and camel fetuses by real-time PCR, respectively. The sensitivity and specificity of real-time PCR obtained 100% and 100%. Statistical analysis showed significant differences (p<0.01) between B. abortus and B. melitensis that were detected in abomasal contents of aborted bovine, ovine, caprine, buffalo and camelid fetuses and between presences of Brucella spp. in bovine with caprine, buffalo and camel aborted fetuses (p<0.05). The CT values obtained from real-time PCR had significant differences between aborted bovine, ovine, caprine, buffalo and camel fetuses for presence of B. abortus and B. melitensis. Results showed that the real-time PCR is considerably faster than current standard methods for isolation and segregation of Brucella spp. Keywords : aborted fetuses, Brucella abortus, Brucella melitensis, conventional PCR, real-time PCR

• Publication year

  2012

• Venue

  Wetchasan sattawaphaet = The Thai journal of veterinary medicine

• Publication date

  2012-03-01

• Fields of study

  Agricultural And Food Sciences, Biology

• Identifiers
  DOI 10.56808/2985-1130.2361
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar

• No claims are published for this paper.

• No concepts are published for this paper.

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