AI Article Synopsis
- - A PCR-based test was created to detect Salmonella by amplifying a specific 93-bp gene fragment involved in the bacterium's invasion of HeLa cells, particularly focusing on Salmonella ser Typhi strain Ty2.
- - The test uses a non-radioactive sandwich hybridization method in microtiter plates, incorporating specific oligonucleotides and a detection system involving avidin linked to alkaline phosphatase.
- - This method is efficient and adaptable for large-scale testing, as it requires only a thermal cycler and a standard microtiter plate reader.
Article Abstract
A polymerase chain reaction (PCR)-based test was developed for the detection of Salmonella. One pair of oligonucleotide primers was designed to amplify a 93-bp fragment of a gene required for the invasion of HeLa cells by Salmonella ser Typhi strain Ty2. The amplified product was analysed by non-radioactive sandwich hybridisation in microtiter plates using two oligonucleotides. The capture oligonucleotide was covalently linked onto animated wells of microtiter plates. The detection oligonucleotide was labelled with biotine. The hybrid molecules were detected by avidine conjugated with alkaline phosphatase and chromogenic substrate. The described combination of microplate sandwich hybridisation and PCR seems to be a suitable method for rapid detection of Salmonella subspecies I. It only requires a thermal cycler and a conventional microtiter reader, and can be readily done on a large scale.
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| http://dx.doi.org/10.1111/j.1574-695X.1995.tb00039.x | DOI Listing |
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