Manzulli Viviana, Rondinone Valeria, Serrecchia Luigina, Petrella Antonio, Scaltrito Domenico, Marino Leonardo, Pace Lorenzo, Luigia Prencipe Maria, Cipolletta Dora, Nitti Mauro, Fasanella Antonio, Poppa Elena, Galante Domenico
Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, 71121 Foggia, Italy.
Azienda Sanitaria Locale Avellino, 83100 Avellino, Italy.
J Vet Res. 2022 Dec 18;66(4):559-563. doi: 10.2478/jvetres-2022-0062. eCollection 2022 Dec.
Brucellosis is a widespread zoonosis of great economic importance for livestock farming in many areas of the world. It is a highly infectious disease which is diagnosed using conventional serological and microbiological methods. The aim of this study was to assess the efficiency of a specific real-time PCR in combination with broth cultivation in detecting spp. in organs of infected cattle, in order to compare the sensitivity of the two approaches and the time needed in them until a correct diagnosis is made.
We examined 67 organs collected from 10 cattle slaughtered following a brucellosis outbreak which occurred in February 2016 in southern Italy. The research was carried out by enrichment broth cultivations in combination with a real-time PCR every week for six weeks.
strains were isolated by cultivation from 44 enrichment broths of organs. All isolates were later identified as by real-time PCR. Using this method in combination with cultivation made it possible to identify the same percentage of infected animals faster than by cultivation alone. Moreover, the same diagnostic results were obtained, on average two weeks before they would have been using only cultivation. In almost all cases, was detected by real-time PCR after the first week of cultivation in pre-enrichment broth, while the bacterial growth was evident usually after 2 or 3 weeks.
Real-time PCR has allowed results to be obtained faster than in the classical microbiological method, reducing the response times to identify positive animals by half.
布鲁氏菌病是一种广泛传播的人畜共患病，对世界许多地区的畜牧业具有重大经济意义。它是一种高度传染性疾病，可通过传统血清学和微生物学方法进行诊断。本研究的目的是评估特异性实时荧光定量聚合酶链反应（PCR）结合肉汤培养法在检测感染牛器官中布鲁氏菌属细菌的效率，以比较这两种方法的灵敏度以及做出正确诊断所需的时间。
我们检查了从2016年2月意大利南部发生布鲁氏菌病疫情后屠宰的10头牛身上采集的67个器官。研究通过富集肉汤培养结合实时荧光定量PCR进行，为期六周，每周进行一次。
从44个器官的富集肉汤培养物中分离出布鲁氏菌菌株。所有分离株随后通过实时荧光定量PCR鉴定为布鲁氏菌。将这种方法与培养法结合使用，能够比单独使用培养法更快地鉴定出相同比例的感染动物。此外，获得了相同的诊断结果，平均比仅使用培养法提前两周。在几乎所有情况下，在预富集肉汤培养第一周后通过实时荧光定量PCR检测到布鲁氏菌，而细菌生长通常在2或3周后明显可见。
实时荧光定量PCR比经典微生物学方法能更快获得结果，将鉴定阳性动物的响应时间缩短了一半。
