scholarly journals Detection of Brucella abortus in Bovine Milk by Polymerase Chain Reaction

2010 ◽  
Vol 79 (2) ◽  
pp. 277-280 ◽  
Author(s):  
Ayman Al-Mariri ◽  
Nermeen Haj-Mahmoud

This study compares milk ring test and three different polymerase chain reaction techniques (direct DNA extraction by column purification system, alkaline DNA extraction, and filtrated milk), in order to identifyBrucella abortusinfection in bovine milk. Milk ring test sensitivity and specificity were 72% and 80%, respectively. While specificity of the three polymerase chain reaction techniques was 100%; sensitivity was 92%, 88% and 100%, respectively, for the three polymerase chain reaction procedures. We conclude that the filtered animal’s milk polymerase chain reaction is the best procedure to make the diagnosis ofB. abortusinfections.

Author(s):  
Susan M. Noor ◽  
Pratiwi Sudharmono ◽  
Asmarani Kusumawati ◽  
Anis Karuniawati

Penelitian ini bertujuan mendeteksi brucelosis pada sampel susu sapi dengan uji polymerase chain reaction (PCR) dan membandingkan tingkatsensitivitas dan spesifisitasnya dengan metode milk ring test (MRT). Sebanyak 24 sampel susu sapi yang dikoleksi secara aseptik dari lapang diuji PCR dan MRT. Hasil pengujian menunjukkan bahwa 79,17% (19/24) sampel susu positif brucelosis dengan uji PCR dan 83,33% (20/24) dengan uji MRT. Sensitivitas dan spesifisitas PCR mendeteksi brucelosis masing-masing sebesar 75 dan 100% dibandingkan dengan uji MRT.


Author(s):  
Dwiyitno Dwiyitno ◽  
Stefan Hoffman ◽  
Koen Parmentier ◽  
Chris Van Keer

Fish and seafood products has been commonly targeted for fraudulent activities. For that reason, authentication of fish and seafood products is important to protect consumers from fraudulent and adulteration practices, as well as to implement traceability regulation. From the viewpoint of food safety, authenticity is beneficial to protect public from serious food poisoning incidents, such as due to ingestion of toxic species. Since DNA based identification depends on the nucleic acid polymerase chain reaction (PCR), the quantity and quality/purity of DNA will contribute significantly to the species authentication. In the present study, different DNA extraction and purification methods (3 classical methods and one commercial kit) were compared to produce the better isolated DNA for PCR amplification. Additionally, different methods for the estimation of DNA concentration and purity which is essential for PCR amplification efficiency were also evaluated. The result showed that classical DNA extraction methods (based on TNES-Urea) yielded a higher amount of DNA (11.30-323.60 ng/g tissue) in comparison to commercial kit/Wizard Promega (5.70-83.45 ng/g tissue). Based on the purity of DNA extract (A260/280), classical DNA extraction method produced relatively similar on DNA quality to the commercial kit (1.79-2.12). Interestingly, all classical methods produced DNA with A260/280 ratio of more than 2.00 on the blue mussel, in contrast with commercial kit. The commercial kit also produced better quality of DNA compared to the classical methods, showing the higher efficiency in PCR amplification. NanoDrop is promising as cheap, robust and safe UV-spectrophotometer method for DNA quantification, as well as the purity evaluation.Keywords: seafood authenticity, DNA isolation, polymerase chain reaction, NanoDrop, Picogreen


2016 ◽  
Vol 40 (1) ◽  
pp. 5-8
Author(s):  
Bashar Sadeq Noomy

      The aim of this study is to determine the sensitivity of bacterial culture technique in the detection of Brucella abortus in milk samples of aborted cows. Sixty samples of milk were collected from aborted cows during a period which did not exceed two months after the abortion. All of them were positive for rose bengal test. Results showed that Brucella abortus was isolated from 7 out of 60 (11.6%) from the milk of aborted cows, while PCR test showed that 32 out of 60 (53.3%) milk sample contained Brucella abortus. The specificity of culture techniques was 10%, but its sensitivity was only 21.8%. Beside the cautions in dealing with live Brucella abortus (as culture), it is also less sensitive than PCR, though it is better to use PCR technique in the diagnosis of brucellosis in aborted cows milk.


Author(s):  
Fernanda Pinto-Ferreira ◽  
Jonatan Batista Reis ◽  
Aline Ticiani Pereira Paschoal ◽  
Letícia Santos Balbino ◽  
Amanda Bertão-Santos ◽  
...  

Abstract The consumption of vegetables has increased in recent years due to the search for a healthier diet that is rich in fiber and has fewer calories. To assess the parasitic contamination of lettuce sold in markets, a survey of parasites was carried out from a supermarket chain in the city of Londrina, Paraná. A total of thirty samples of lettuce were purchased in the ten markets visited, three in each, of which ten were conventionally cultivated, ten were hydroponically cultivated, and ten were organically cultivated. All samples were analyzed using the sedimentation methods of Hoffman, Pons and Janer and the fluctuation method of Faust and colleagues and Willis with adaptations. In addition, the samples were subjected to DNA extraction by a commercial kit and polymerase chain reaction to detect Toxoplasma gondii, Cryptosporidium spp. and Giardia spp., which are protozoa that cause food and waterborne parasitic outbreaks. All samples were negative for sedimentation and flotation techniques. One of the hydroponically cultivated samples was positive for T. gondii. The results demonstrate the risk of curly lettuce contamination from hydroponic cultivation and the need for proper cleaning of these foods before consumption.


Author(s):  
Hayder Mohammad Al-Rammahi ◽  
Abdulameer Abed Hatem ◽  
Asaad Chasib Al-Atabi

This study was designed to detect equine piroplasmosis using the molecular technique in Al-Najaf province during the season that showed an increment in tick activities. Blood samples were collected from 110 horses with more than two signs of piroplasmosis. After DNA extraction, the product was examined by a polymerase chain reaction to amplify 18SrRNA. The results showed that the overall percentage of equine theileriosis was 38.18%. According to gender, the percentage of infection was 43.48% and 29.27% in females and males, respectively. Significant variations appeared between infected horses according to age, and the percentage of infection was 50% and 35.22% in less than 2 years and more than 2 years age, respectively. Moreover, the percentage of infection was 62.5% and 19.35% in animals with and without acariasis, respectively. Significant variations were also seen in equine theileriosis according to geographical areas, and the higher percentage was reported in Hera district (60.87%), while the lowest percentage was in the center of Al-Najaf (21.43%). This difference may be due to different distribution of vector of disease (tick), which may be the availability of the suitable weather that helped in the multiplication of the intermediate vectors. In conclusion, this study proved the variations in the occurrences of equine piroplasmosis according to gender, age, and geographical areas.


Sign in / Sign up

Export Citation Format

Share Document