The main methodological problems of multi-criteria analysis of the effectiveness of parasitological examination

The present scientific research is aimed at systematizing general approaches to the choice of multiplex PCR in clinical and sanitary parasitology. Our data are based on the need to focus the laboratory service, which carries out parasitological control of environmental safety, on the development and organization of multi-complex studies, which, based on a provide the most comprehensive data for a factor analysis and multi-criteria assessment of the parasitological situation in the country.
The purpose of the study: Conducting an analysis of current trends and our own multicriteria data for evaluating the effectiveness of laboratory tests based on PCR analysis in parasitology.
Materials and methods. The analysis of modern literature data from peer-reviewed sources (PubMed) has been carried out to substantiate the scientific purpose of the study. A series of experimental studies of two sets of multiplex PCR (qPCR) tests for the detection of 7 helminth species contained in 192 model samples was carried out.
Research results. The methodological limitations and advantages of using microscopic and multimplex PCR studies are determined. The limits of detection of DNA from helminth eggs and fragments of imago tissues of round and flat helminths stored for a long time in preservative, as well as in freshly isolated biomaterial, have been established at the level of 101-103 in 100 µl and the diagnostic sensitivity of PCR tests at the level of 70,0-90,0 %.
Conclusion. It is proposed to develop an innovative form of multi-criteria assessment of environmental parasitic safety based on the widespread use of multiplex PCR (qPCR) tests, and in the future, with the introduction of digital PCR (dPCR) to simultaneously determine the qualitative and quantitative characteristics of pathogenic contaminants of different types when testing uniform samples. Based on the data obtained, the necessity of active development of PCR technologies and their wide application in parasitology in the laboratory service of state control bodies is substantiated.

1. Kaisar M.M.M., Brienen E.A.T., Djuardi Y., Sartono E., Yazdanbakhsh M., Verweij J.J., Supali T., VAN Lieshout L. Improved diagnosis of Trichuris trichiura by using a beadbeating procedure on ethanol preserved stool samples prior to DNA isolation and the performance of multiplex realtime PCR for intestinal parasites // Parasitology. 2017; 144(7): 965-974. DOI: 10.1017/S0031182017000129.

2. Pomari E., Perandin F., La Marca G., Bisofi Z. Improved detection of DNA Schistosoma haematobium from eggs extracted by bead beating in urine // Parasitol Res. 2019; 118(2): 683-686. doi: 10.1007/s00436-018-6137-7.

3. Pomari E., Kabelli K., Perandin F., Bisofi Z. Digital PCR: a new technology for the diagnosis of parasitic infections // Clin Microbiol Infect. 2019; 25: 1510–6. DOI: 10.1016/j.cmi.2019.06.009.

4. PohlG.,ShihIeM.Principleandapplications of digital PCR // Expert Rev Mol Diagn. 2004; 4(1): 41-7. DOI: 10.1586/14737159.4.1.41.

5. Amoah I.D., Singh G., Stenström T.A., Reddy P. Detection and quantification of soil-transmitted helminths in environmental samples: A review of current state-of-the-art and future perspectives // Trop. 2017; 169: 187-201. DOI: 10.1016/j.actatropica.2017.02.014.

6. Dunn J.C., Turner H.C., Tun A., Anderson R.M. Epidemiological surveys of, and research on, soil-transmitted helminths in Southeast Asia: a systematic review // Parasit Vectors. 2016; 9 :31. DOI: 10.1186/s13071-016-1310.

7. Morozov E.N. Prospects for the application of molecular parasitology methods in monitoring socially significant parasitoses: specialty 03.02.11 "Parasitology": dissertation for the degree of Doctor of Biological Sciences. M, 2016: 189.(in Russian)

8. Ayana M., Cools P., Mekonnen Z., Biruksew A., Dana D., Rashwan N., Prichard R., Vlaminck J., Verweij J.J., Levecke B. Comparison of four DNA extraction and three preservation protocols for the molecular detection and quantification of soiltransmitted helminths in stool // PLoS Negl Trop Dis. 2019; 13(10): e0007778. DOI: 10.1371/journal.pntd.0007778.

9. Manuel M., Amato H.K., Pilotte N., Chieng B., Araka S.B., Siko J.E.E., Harris M., Nadimpalli M.L., Janagaraj V., Houngbegnon P., Rajendiran R., Thamburaj J., Kaliappan S.P., Sirois A.R., Walch G., Oswald W.E., Asbjornsdottir K.H., Galagan S.R., Walson J.L., Williams S.A., Luty A.J.F., Njenga S.M., Ibikounlé M., Ajjampur S.S.R., Pickering A.J. Soil surveillance for monitoring soiltransmitted helminths: Method development and field testing in three countries // PLoS Negl Trop Dis. 2024; 18(9): e0012416. DOI: 10.1371/journal.pntd.0012416.

10. Phuphisut O., Yoonuan T., Sanguankiat S., Chaisiri K., Maipanich W., Pubampen S., Komalamisra C., Adisakwattana P. Triplex polymerase chain reaction assay for detection of major soil-transmitted helminths, Ascaris lumbricoides, Trichuris trichiura, Necator americanus, in fecal samples // Southeast Asian J Trop Med Public Health. 2014; 45(2): 267-75.

11. Hasegawa M., Pilotte N., Kikuchi M., Means A.R., Papaiakovou M., Gonzalez A.M., Maasch J.R.M.A., Ikuno H., Sunahara T., Ásbjörnsdóttir K.H., Walson J.L., Williams S.A., Hamano S. What does soil-transmitted helminth elimination look like? Results from a targeted molecular detection survey in Japan // Parasit Vectors. 2020; 13(1): 6. DOI: 10.1186/s13071-019-3875-z.

12. Basic laboratory methods in medical parasitology. 1. Parasitology - laboratory manuals. WHO. Geneva. 1991.

13. Sergiev V.P. The registered and true prevalence of parasitic diseases // Medical parasitology and parasitic diseases. 1991; №2: 3-5. (in Russian)

14. Onishchenko G.G. The incidence of parasitic diseases in the Russian Federation and the main activities for its stabilization // Medical parasitology and parasitic diseases. 2002; 4: 3-10. (in Russian)

15. Tverdokhlebova T.I. Trichinellosis in the South of Russia: Epidemiology, Diagnostics and Prevention in Modern Socio-Economic Conditions: Specialty 03.02.11 "Parasitology": Dissertation for the Degree of Doctor of Biological Sciences. Moscow, 2007: 282

16. Stepanova T.F., Kornchaev A.S. Approaches to improving the system of surveillance and management of the epidemic process of parasitic diseases: monograph; Federal Budgetary Scientific Institution Tyumen Scientific Research Institute of Regional Infectious Pathology of the Federal Service for Surveillance on Consumer Rights Protection and Human Wellbeing. Tyumen: Publishing House of Tyumen State University, 2012: 147.