Identification of Brucella melitensis from camel’s blood by vitek2 and real time polymerase chain reaction

Kavitha Manivannan, Malathi Ramasamy, Uma Sundaresan, Samar M. Moustafa, Sherloumay, Safna Mariyam
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Abstract

Introduction and aim. Brucellosis is a zoonotic disease. Experimental clinical and laboratory diagnosis is still facing problems in identifying the organism. The present study will diagnose a Brucella infection in camel blood in Qatar using serological assays. Isolation and identification were performed on a camel blood sample. Brucella in bacterial isolates was determined by real-time polymerase chain reaction (RT-PCR) as a gold standard test. Material and methods. A total of 220 samples, 200 random serum samples, and 20 EDTA blood samples were selected among the above-mentioned random samples, and 20 serum samples from camel handlers were collected from Al Shahaniya prov ince, Qatar. The Rose Bengal test (RBT), buffered antigen plate agglutination test (BAPAT), and enzyme linked immunosorbent assay (cELISA) for the monoclonal antibody in serum samples were performed using commercially available kits. For the molecular detection of Brucella, conventional PCR and real-time PCR (GPS kit) were used for the genus-specific insertion sequence IS711. Brucella melitensis (MICROBOSS Hightech GmbH kit) was used to identify subspecies. Results. The results identified by vitek2 compact (30%) showed B. melitensis in 6 samples out of 20 isolates. Both conventional (66.67%) and RT-PCR (83.33%) analyses supported this, demonstrating the presence of Brucella. These tests also showed that Brucella species were present in Rose Bengal 182/200 (91%), BAPAT 182/200 (91%), and cELISA (90%) 180/200 in camel serum. Conclusion. To conclude, the prevalence of brucellosis in dromedary camels is higher in this region, and as a matter of urgency, measures should be taken to control the disease.
用 vitek2 和实时聚合酶链反应鉴定骆驼血液中的布鲁氏菌
导言和目的。布鲁氏菌病是一种人畜共患病。实验性临床和实验室诊断在确定病原体方面仍面临问题。本研究将利用血清学检测方法诊断卡塔尔骆驼血液中的布鲁氏菌感染。对骆驼血液样本进行了分离和鉴定。细菌分离物中的布鲁氏菌是通过实时聚合酶链反应(RT-PCR)作为金标准检测来确定的。材料和方法从卡塔尔 Al Shahaniya 省采集了 220 份样本,其中 200 份为随机血清样本,20 份为 EDTA 血液样本,20 份为骆驼饲养者的血清样本。使用市售试剂盒对血清样本中的单克隆抗体进行了玫瑰红试验(RBT)、缓冲抗原平板凝集试验(BAPAT)和酶联免疫吸附试验(cELISA)。在布鲁氏菌的分子检测中,使用了传统 PCR 和实时 PCR(GPS 试剂盒)检测属特异性插入序列 IS711。梅里特布鲁氏菌(MICROBOSS Hightech GmbH 试剂盒)用于鉴定亚种。结果。通过 vitek2 compact(30%)鉴定的结果显示,在 20 个分离样本中,有 6 个样本含有梅里特氏布鲁菌。传统分析(66.67%)和 RT-PCR 分析(83.33%)都证实了这一点,证明了布鲁氏菌的存在。这些检测还显示,在骆驼血清中,孟加拉玫瑰菌 182/200 (91%)、BAPAT 182/200 (91%)和 cELISA 180/200 (90%)均检出布鲁氏菌。结论总之,该地区单峰骆驼的布鲁氏菌病发病率较高,当务之急是采取措施控制该疾病。
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