The Use of Oral Fluid Samples Spotted on Filter Paper for the Detection of Measles Virus Using Nested RT‐PCR

Abstract

Measles is the leading cause of death in infants, although a vaccine is available for its prevention. At this stage of measles elimination and eradication, it is so important to confirm clinically diagnosed measles cases in the laboratory but, developing countries have troubles in collecting and maintaining the cold chain of the specimens while transporting them to the laboratories. Therefore, filter papers are good candidates for simplification of specimen collection and transportation. In this research, the effects of the temperature, at which the dried specimens were kept, and the time duration the dried specimens were kept before being tested, were studied. Since there were not enough patients' oral fluid samples available, a nested reverse transcriptase PCR (RT-PCR) that detected measles virus (MV) from dried filter papers was set up using MV infected cells diluted in sterile phosphate-buffered saline (PBS). Dried specimens were stored at -25°C, 4°C, and room temperature for 1 day, 1, 2, and 3 weeks before being tested. This method was then applied to filter paper oral fluids collected from nine clinically diagnosed measles patients in Iran in 2010 which were tested after being kept at room temperature for 1 day, 1 and 3 weeks after preparation. The results showed that dried oral fluids on filter papers are reliable specimens for the detection of MV RNA using nested RT-PCR, but the nested RT-PCR results of low titer viruses dried onto filter papers are not reproducible and reliable.