High-Resolution Melting Analysis in Comparison with Microscopic Method: An Experimental Study to Diagnosis of Plasmodium Species Infections in Human
-
Mahya ALLAHMORADI
,
Afsaneh MOTEVALLI HAGHI
,
Mehdi NATEGHPOUR
,
Mehdi MOHEBALI
,
Ahmad RAEISI
,
Ahmad HOSSEINI SAFA
,
Sina MOHTASEBI
,
Mohammad Javad ABBASZADEH AFSHAR
Abstract
Background: Among the human parasitic diseases, malaria is the main cause of morbidity and mortality. To prevent the high mortality and tracking malaria elimination efforts, a prompt and sensitive diagnosis is essential. This study aimed to compare High-Resolution Melting (HRM) and microscopic methods to diagnose Plasmodium falciparum and P. vivax.
Methods: Eighty-one blood samples were collected from patients with clinical symptoms who were suspect to malaria in Chabahar district, southeastern Iran and also, from those who were referred to Malaria National Laboratory in the Tehran University of Medical Sciences, Tehran, Iran. Microscopic examination and HRM method were used to the diagnosis of Plasmodium parasites simultaneously.
Results: Microscopic results revealed 45 positive cases (12 P. falciparum and 33 P. vivax) out of 81 collected samples while according to HRM analysis results 11 and 33 samples were identified as P. falciparum and P. vivax, respectively. HRM analysis also revealed 1 mixed infection of P. falciparum and P. malariae.
Conclusion: HRM analysis provides a promising mean for simultaneous detection and discrimination of the Plasmodium spp. especially in mixed infection cases.
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13. Farcas GA, Zhong KJ, Lovegrove FE, et al. Evaluation of the Binax NOW® ICT test versus polymerase chain reaction and microscopy for the detection of malaria in returned travelers. Am J Trop Med Hyg. 2003;69(6):589-92.
14. Snounou G, Viriyakosol S, Jarra W, et al. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol.1993;58(2):283-92.
15. Mixson-Hayden T, Lucchi NW, Udhayakumar V. Evaluation of three PCR-based diagnostic assays for detecting mixed Plasmodium infection. BMC Res Notes. 2010;3:88.
16. Chew CH, Lim YAL, Lee PC, et al. Hexaplex PCR detection system for identification of five human Plasmodium species with an internal control. J Clin Microbiol. 2012;50(12):4012-9.
17. Andriantsoanirina V, Lascombes V, Ratsimbasoa A, et al. Rapid detection of point mutations in Plasmodium falciparum genes associated with antimalarial drugs resistance by using High-Resolution Melting analysis. J Microbiol Methods. 2009;78(2):165-70.
18. Robinson BS, Monis PT, Dobson PJ. Rapid, sensitive, and discriminating identification of Naegleria spp. by real-time PCR and melting-curve analysis. Applied and Environmental Microbiology. 2006;72(9):5857-63.
19. Talmi-Frank D, Nasereddin A, Schnur LF, et al. Detection and identification of old world Leishmania by high resolution melt analysis. PLoS Negl Trop Dis. 2010;4(1):e581.
20. Hosseini-Safa A, Rokni Mb, Mosawi Sh, et al. High-Resolution Melting Analysis as an Appropriate Method to Differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health. 2019;48(3):501-7.
21. Safa AH, Harandi MF, Tajaddini M, et al. Rapid identification of Echinococcus granulosus and E. canadensis using high-resolution melting (HRM) analysis by focusing on single nucleotide polymorphism. Jpn J Infect Dis.2015; 69(4):300-5.
22. Brown AE, Kain KC, Pipithkul J, et al. Demonstration by the polymerase chain reaction of mixed Plasmodium falciparum and P. vivax infections undetected by conventional microscopy. Trans R Soc Trop Med Hyg. 1992;86(6):609-612.
23. Zakeri S, Najafabadi ST, Zare A, et al. Detection of malaria parasites by nested PCR in south-eastern, Iran: evidence of highly mixed infections in Chahbahar district. Malar J. 2002;1:2.
2. Chua KH, Lim SC, Ng CC, et al. Development of high resolution melting analysis for the diagnosis of human malaria. Sci Rep. 2015;5:15671..
3. Perlmann P, Troye-Blomberg M. Malaria blood-stage infection and its control by the immune system. Folia Biol (Praha). 2000;46(6):210-8..
4. Edrissian G. Malaria in Iran: Past and present situation. Iran J Parasitol .1(1):1-14.
5. Abba K, Deeks JJ, Olliaro PL, et al. Rapid diagnostic tests for diagnosing uncomplicated P. falciparum malaria in endemic countries. Cochrane Database Syst Rev. 2011:2011(7).
6. Pasay C, Arlian L, Morgan M, et al. High‐resolution melt analysis for the detection of a mutation associated with permethrin resistance in a population of scabies mites. Med Vet Entomol. 2008;22(1):82-8.
7. Espy M, Uhl J, Sloan L, et al. Real-time PCR in clinical microbiology: applications for routine laboratory testing. Clin Microbiol Rev. 2006;19(1):165-256.
8. Organization WH. Basic malaria microscopy: Part I. Learner's guide. Basic malaria microscopy: Part I Learner's guide. 2010(Ed. 2).
9. Higuera SL, Guhl F, Ramírez JD. Identification of Trypanosoma cruzi Discrete Typing Units (DTUs) through the implementation of a High-Resolution Melting (HRM) genotyping assay. Parasit Vectors. 2013;6:112..
10. Hemami MR, Sari AA, Raeisi A, et al. Malaria elimination in Iran, importance and challenges. Int J Prev Med. 2013;4(1):88.-94.
11. Kipanga PN, Omondi D, Mireji PO, et al. High-resolution melting analysis reveals low Plasmodium parasitaemia infections among microscopically negative febrile patients in western Kenya. Malar J. 2014;13:429.
12. Ehtesham R, Fazaeli A, Raeisi A, et al. Detection of mixed-species infections of Plasmodium falciparum and Plasmodium vivax by nested PCR and rapid diagnostic tests in southeastern Iran. Am J Trop Med Hyg. 2015;93(1): 181-5.
13. Farcas GA, Zhong KJ, Lovegrove FE, et al. Evaluation of the Binax NOW® ICT test versus polymerase chain reaction and microscopy for the detection of malaria in returned travelers. Am J Trop Med Hyg. 2003;69(6):589-92.
14. Snounou G, Viriyakosol S, Jarra W, et al. Identification of the four human malaria parasite species in field samples by the polymerase chain reaction and detection of a high prevalence of mixed infections. Mol Biochem Parasitol.1993;58(2):283-92.
15. Mixson-Hayden T, Lucchi NW, Udhayakumar V. Evaluation of three PCR-based diagnostic assays for detecting mixed Plasmodium infection. BMC Res Notes. 2010;3:88.
16. Chew CH, Lim YAL, Lee PC, et al. Hexaplex PCR detection system for identification of five human Plasmodium species with an internal control. J Clin Microbiol. 2012;50(12):4012-9.
17. Andriantsoanirina V, Lascombes V, Ratsimbasoa A, et al. Rapid detection of point mutations in Plasmodium falciparum genes associated with antimalarial drugs resistance by using High-Resolution Melting analysis. J Microbiol Methods. 2009;78(2):165-70.
18. Robinson BS, Monis PT, Dobson PJ. Rapid, sensitive, and discriminating identification of Naegleria spp. by real-time PCR and melting-curve analysis. Applied and Environmental Microbiology. 2006;72(9):5857-63.
19. Talmi-Frank D, Nasereddin A, Schnur LF, et al. Detection and identification of old world Leishmania by high resolution melt analysis. PLoS Negl Trop Dis. 2010;4(1):e581.
20. Hosseini-Safa A, Rokni Mb, Mosawi Sh, et al. High-Resolution Melting Analysis as an Appropriate Method to Differentiate between Fasciola hepatica and F. gigantica. Iran J Public Health. 2019;48(3):501-7.
21. Safa AH, Harandi MF, Tajaddini M, et al. Rapid identification of Echinococcus granulosus and E. canadensis using high-resolution melting (HRM) analysis by focusing on single nucleotide polymorphism. Jpn J Infect Dis.2015; 69(4):300-5.
22. Brown AE, Kain KC, Pipithkul J, et al. Demonstration by the polymerase chain reaction of mixed Plasmodium falciparum and P. vivax infections undetected by conventional microscopy. Trans R Soc Trop Med Hyg. 1992;86(6):609-612.
23. Zakeri S, Najafabadi ST, Zare A, et al. Detection of malaria parasites by nested PCR in south-eastern, Iran: evidence of highly mixed infections in Chahbahar district. Malar J. 2002;1:2.
| Files | ||
| Issue | Vol 15 No 3 (2020) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijpa.v15i3.4205 | |
| Keywords | ||
| Malaria Diagnosis Microscopy Plasmodium | ||
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How to Cite
1.
ALLAHMORADI M, MOTEVALLI HAGHI A, NATEGHPOUR M, MOHEBALI M, RAEISI A, HOSSEINI SAFA A, MOHTASEBI S, ABBASZADEH AFSHAR MJ. High-Resolution Melting Analysis in Comparison with Microscopic Method: An Experimental Study to Diagnosis of Plasmodium Species Infections in Human. Iran J Parasitol. 2020;15(3):403-410.
