Quick test finds signs of diarrhoeal disease

Bioengineers at Rice University and the University of Texas Medical Branch (UTMB) at Galveston have developed a simple, highly sensitive and efficient test for the diarrhoeal disease cryptosporidiosis that could have great impact on global health.
Results from the diagnostic developed by the lab of Rice bioengineer Rebecca Richards-Kortum are read from a paper strip that resembles a pregnancy test. Lines on the strip tell whether samples taken from the stool of a patient contain genetic DNA from the parasite that causes the disease.
‘Diarrhoeal illness is a leading cause of global mortality and morbidity,’ said Richards-Kortum, director of the Rice 360˚: Institute for Global Health Technologies. ‘Parasites such as cryptosporidium are more common causes of prolonged diarrhoea. Current laboratory tests are not sensitive, are time-consuming and require days before results are available. A rapid, affordable, accurate point-of-care test could greatly enhance care for the underserved populations who are most affected by parasites that cause diarrhoeal illness.’
A. Clinton White, director of the Infectious Disease Division at UTMB, asked Richards-Kortum to help develop a diagnostic test for the parasite. ‘I’ve been working with cryptosporidium for more than 20 years, so I wanted to combine her expertise in diagnosis with our clinical interest,’ he said. ‘Recent studies in Africa and South Asia by people using sophisticated techniques show this organism is a very common, under-appreciated cause of diarrhoeal disease in under-resourced countries.’
Current specialized tests that depend on microscopic or fluorescent analysis of stool samples or polymerase chain reactions (PCR) that amplify pathogen DNA are considered impractical for deployment in developing countries because of the need for expensive equipment and/or the electricity to operate it.
The Rice test depends on recent developments in a recombinase polymerase amplification (RPA) technique that gives similar ‘gold standard’ results to PCR but operates between room and body temperatures. In Rice’s experiments, samples were prepared with a commercial chemical kit that releases all the DNA and RNA in the small amount of stool tested. The purified nucleic acids are then combined with RPA primers and enzymes tuned to amplify the pathogen of interest, Crannell said.
‘If the pathogen DNA is present, these primers will amplify it billions of times to a level that we can easily detect,’ he said. The sample is then flowed over the detection strip, which provides a positive or negative result.
The RPA enzymes are stable in their dried form and can be safely stored at the point of care without refrigeration for up to a year, he said.
While current tests might catch the disease in samples with thousands of the pathogens, the Rice technique detects the presence of very few – even one – parasite in a sample. In their experiments, the researchers reported the presence or absence of the disease was correctly identified in 27 of 28 infected and control-group mice and all 21 humans whose stool was tested. Rice University