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IDENTIFICATION OF Aspergillus flavus FROM INTERNAL TRANSCRIBED SPACER REGIONS 1 AND 2 USING POLYMERASE CHAIN REACTION

Aspergillus flavus is identified as one of the most frequent causes of invasive aspergillosis in humans and animals. This organism is responsible for the elaboration of the toxin in the feed and the toxin is known as aflatoxin. There are various methods available to analyze aflatoxin in foods and feeds which can be classified as rapid, qualitative and quantitative methods. The aim of the project is to develop a rapid detection test kit for aflatoxin using Enzyme-labelled immunoassay technique. For this preliminary study, the primary goal is to produce aflatoxin antigen. In order to produce it, identification of the species available is necessary. Previously, this is usually done by morphological characteristics. Nowadays, a number of molecular, immunological and biochemical methods are available. In this study, the identification of A. flavus species, which is the main aflatoxin producer, was based on internal transcribed spacer (ITS) region using PCR methods. Previous studies indicated that, the ITS regions are located between the 18S and 28S rRNA genes and offer distinct advantages over other molecular targets including increased sensitivity due to the existence of approximately 100 copies per genome. By using this method, the identification of A. flavus was successfully conducted. In future, the toxin produced from this study can be used for the development of ELISA test kit for rapid aflatoxin detection. A quick and easy method for detecting aflatoxin in feed will help farmers to diagnose aflatoxicosis and treat it accordingly to minimise losses due to morbidity and mortality.

Keywords: Aspergillus, ITS Region 1 and 2, aspergillosis

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NORLINDAWATI, P.1, BOHARI, J.1, NAJAMUDDIN, Y.1, RAMLAN, M.1, NORAZURA, H.1, UMMUL HANAN, M.2, NOR FARAH SHAMIRA, H.2

1Veterinary Research Institute, 59 Jalan Sultan Azlan Shah, 31400 Ipoh, Perak

2Universiti Teknologi MARA, Shah Alam


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