Naslov
A pilot study applicability of standardized modified method of dry (throat/buccal) swabs in PCR diagnosis of enteroviral infections

Autori
Borsanyiova, Maria ; Sarmirova, Sona ; Puljak, Livia ; Jeličić Kadić, Antonia ; Borić, Matija ; Vari, Sandor G ; Bopegamage, Shubhada

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Skup
Bridges in Life Sciences 9th Annual Scientific Conference

Mjesto i datum
Split, Hrvatska, 27.05.2014. - 01.06.2014

Vrsta sudjelovanja
Predavanje

Vrsta recenzije
Međunarodna recenzija

Ključne riječi
enteroviruses; throat swabs; buccal swabs; diagnosis; PCR

Sažetak
Introduction: Enteroviruses (EVs) spread via the fecal-oral route. They infect the oro-pharyngeal mucosal cells and tonsils. After replicating at these sites, they pass on to the alimentary tract. We standardized modified methodology of dry throat swabs for EV diagnosis by PCR. Among the commonly collected material for diagnosis besides the stool and blood samples are throat swabs, used for isolations and identifications and PCR diagnosis. The EVs are relatively stable. Commonly for diagnosis the throat swabs are collected in transport medium. We recently standardized the dry swab method and checked the applicability in 10 random volunteers. The rationale of the present study is to standardize the sample collection for diagnosis/screening of EV infections, and to make the sample collection as convenient the patient, staff, and transportation. Methods: Thirty seven volunteers (RECOOP HST Association 4th TriNet Meeting participants) from Croatia, Czech Republic, Hungary, Poland, Romania, Slovak Republic, Ukraine and USA attended the pilot screening study. The sample processed in the Enterovirus laboratory, Slovak Medical University (SMU), Bratislava. As buccal swabs are more convenient to obtain, we collected buccal swabs (two swabs per person for diagnosis of EVs) were from volunteers. Swab-A inserted in sterile test tube and immediately frozen at -80°C and transported from Split to Bratislava on dry ice. Swab-B allowed drying for 1 hour at ambient temperature and processed (vortexed in RNAse-free water) after that the Swab B samples were transferred from Split to Bratislava on dry ice. The resulting suspension was frozen at -80°C. RNA isolation and reverse transcriptase and nested PCR were performed in Bratislava at SMU. Results: Viral RNA was detectable in 6/37 (16.22%) volunteers as compared to 7/10 (70%) throat swabs collected during standardization. In the present study positive samples were random in different countries. Evaluation on terms of intensity of the PCR reaction showed that Swab-A (immediately frozen at -80°C) methodology was more effective for screening or diagnosis. Conclusion: In the samples collected during the RECOOP HST Association meeting, we did not observe a high incidence of EV infections. We conclude the dry swab- method is efficient. We recommend that prior to transport to the Virological laboratory 1.the swabs should be dried at ambient temperatures (if in absence of transport medium), inserted into the sterile containers, 2.frozen at -20°C to -80°C or storage at 4°C depending on availability of freezers and quickness of transport to the Virological laboratory, 3. Considering the viral pathogenesis and our comparative results of the throat swabs (without ruling out the possibility that the throat swabs could have been collected when the virus was endemic) vs. buccal swabs we suggest the throat swabs to be the choice for collection of samples though they maybe less convenient for the patient and staff.

Izvorni jezik
Engleski

Znanstvena područja
Temeljne medicinske znanosti

POVEZANOST RADA