Application of High-Resolution Melting (HRM) Technique Towards the Detection of Asymptomatic Malaria in a Malaria Endemic Area of Southeastern Iran Under Elimination Program
-
Francan Felix Ouma
,
Mehdi Nateghpour
,
Afsaneh Motevalli Haghi
,
Mehdi Mohebali
,
Leila Farivar
,
Ahmad Hosseini-Safa
,
Sayed Hussain Mosawi
Abstract
Background: Asymptomatic malaria, which usually exists in low parasitemia, acts as the Plasmodium species reservoirs contributing towards malaria transmission. This situation hinders malaria elimination programs in endemic areas, thus necessitating an active case detection with a high sensitive method and treatment of cases. This is why we used a High Resolution Melting (HRM) assay to monitor the trend of asymptomatic malaria in a malaria endemic area of Iran which is under elimination program.
Methods: The peripheral blood was sampled from 271 clinically approved non-febrile individuals from a malaria endemic zone of southeastern Iran for asymptomatic malaria prevalence detection by microscopy, Rapid Diagnostic Tests (RDTs) and HRM methods. The HRM assay was done based on the amplification of 18S SSU rRNA gene.
Results: The HRM assay revealed infections from three individuals out of 271 (1.1% asymptomatic malaria prevalence) from the participants, two Iranian natives with Plasmodium vivax infection and one Pakistani immigrant with P. falciparum infection. Neither microscopy nor RDTs detected Plasmodium spp infections from the 271 non-febrile individuals. The nucleotide sequencing analysis of the positive controls used in this study showed a close homology with the reference gene bank sequences of P. falciparum 3D7 (CPO16995.1) and P. vivax Sal-1(UO3079.1).
Conclusion: This study revealed a low frequency of asymptomatic malaria trend within malaria endemic areas of southeastern Iran which are under intense elimination program and also the ability of HRM assay in detecting low Plasmodium spp parasitemia beyond the limits of microscopy and RDTs.
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9. Cooke A, Chiodini PL, Doherty T, Moody AH, Ries J, Pinder M (1999) Compari-son of a parasite lactate dehy¬drogenase-based Immunochromato¬graphic antigen de¬tection assay (opti¬mal) with micros-copy for the detection of malaria para-site in human blood samples. Am J Soc Trop Med Hyg. 60: 173–176.
10. Chua K, Lim S, Ng C, Lee P, Lim YA, Lau, Chai H (2016) Development of High Resolution Melting Analysis for the diagnosis of human malaria. Sci¬en-tific Reports. 5: 15671.
11. Mahende C, Ngasala B, Lusingu J, Yong T, Lushino P, Lemnge M, Mmbando B, Premji Z (2016) Performance of rapid diagnostic test, blood film mi¬croscopy and PCR for diagnosis of malaria infec-tion among febrile chil¬dren from Korog-we district, Tanzania. Malar J. 15: 391–398.
12. Kipanga PN, Omondi D, Mireji PO, Sawa P, Masiga DK, Villinger J (2014) High-resolution analysis reveals low Plasmo-dium parasitaemia infections among mi-cro¬scopically negative fe¬brile patients in western Kenya. Malar J. 13: 429–437.
13. Mangold KA, Manson RU, Koay ES, Ste-phens L, Renger MA, Thomson RB, Pe-terson LR, Kaul KL (2005) Real-time PCR for detection and identifica¬tion of Plasmodium spp. J Clin Micro¬biol. 43: 2435–2440.
14. Canier L, Khim N, Kim S, Sluydts V, Heng S, Dourng D, Eam R, Chy S, Khean C, Loch K, Ken M, Lim H, Siv S, Tho S, Masse-Navette P, Gryseels C, Uk S, Van Roey K, Grietens KP, Sokny M, Thavrin B, Chuor CM, Deubel V, Durnez L, Coosemans M, Menard D (2013) An in¬novative tool for moving malaria PCR detection of parasite reser-voir into the field. Malar J. 12: 405–417.
15. Sargolzaie N, Salehi M, Kiani M, Sakeni M, Hasanzehi A (2011) Malaria epi¬de-miology in Sistan and Baluchestan Prov-ince during April 2008-March 2011, Iran. Zahedan J Res Med Sci. 16: 41–43.
16. Oshaghi MA, Yaghoobi F, Vatandoost H, Abai MR, Akbarzadeh K (2006) Anoph-eles stephensi biological forms, Geo-graph¬ical distribution and malaria transmission in malarious regions of Iran. Pakistan J Biol Sci. 9: 294–298.
17. WHO (2016) Giemsa Staining of Malaria Blood films. Malaria Microscopy Stand-ard Operating Procedure.World Health Organization, Geneva, Swit¬zerland. Avail¬able at: https://apps.who.int/iris/handle/10665/274382
18. Ryan T (2010) Malaria service delivery protocol for sun network. Population services international. Available at: https://silo.tips/download/malaria-ser-vice-delivery-protocol-for-sun-network
19. Bereczky S, Martensson A, Gil JP, Far¬nert A (2005) short report: Rapid DNA ex-traction from archive blood spots on fil-ter paper for genotyping of Plasmo¬dium falciparum. Am J Trop Med Hyg. 72(3): 249–251.
20. Chew CH, Lim YL, Lee PC, Mahmud R, Chua KH (2012) Hexaplex PCR de¬tec-tion system for the identification of five human Plasmodium species with inter-nal controls. J Clin Microbiol. 50(12): 4012–4019.
21. Erali M, Wittwer CT (2010) High Resolu-tion Melting Analysis for gene scan¬ning. PMC. 50: 250–261.
22. Turki H, Raeisi A, Malekzadeh K, Ghan-barnejad A, Zoghi S, Yeryan M, Abedi Nejad M, Mohseni F, Shekari M (2016) Efficiency of Nested-PCR in detecting asymptomatc cases toward malaria elim-ination program in an en¬demic area of Iran. Iran J Parasitol. 10(1): 39–45.
23. Amirshekari MB, Nateghpour M, Raeisi A, Motevalli Haghi A, Farivar L, Edris-sian GhH (2016) Determination of asymp¬tomatic malaria among Af¬ghani and Pakistani immigrants and native population in south of Kerman province, Iran. Iran J Parasitol. 11: 247–252.
24. Nateghpour M, Akbarzadeh K, Farivar L, Amiri A (2011) Detection of asymp¬to-matic malaria among the Afghani immi-grant population in Iranshahr districts of southeastern Iran. Bul Soc Pathol Exot. 104: 321–323.
25. Turki H, Zoghi S, Mehrizi AA, Zakeri S, Raeisi A, Khazan H, Haghdoost AA (2012) Absense of asymptomatic ma-laria infection in endemic area of Ba-shagard District, Hormozgan Prov¬ince, Iran. Ira¬nian J Parasitol. 7: 36–44.
26. Pirahmadi S, Zakeri S, Raeisi A (2017) Ab¬sence of asymptomatic malaria in-fection in a cross-sectional study in Iranshahr District, Iran under elimina-tion pro¬gramme. Iran J Parasitol. 12: 90-100.
27. Gan LS, Loh JP (2010) Rapid identifica-tion of chloroquine and atovaquone drug resistance in Plasmodium falci-parum using High-Resolution Melt Pol-ymerase Chain Reaction. Malar J. 9: 134–139.
28. Hussein EM, Al-Mohammed HI, Hussein AM (2009) Genetic diversity of Dien-tamoeba fragilis isolates of bowel syn-drome patients by High-Resolution Melt¬ing-curve (HRM) analysis. Para¬sitol Res. 105: 1053–1060.
29. Hosseini-Safa AH, Harandi MF, Tajaddini M, Rostami-Nejad M, Mohtashami-Pour M, Pestehchian N (2016) Rapid identi-fication of Echinococcus gran¬ulosus and E. Canadensis using High Resolu-tion Melt¬ing (HRM) analysis by focus-ing on a single nucleotide polymor-phism. J Infect Dis. 69: 300–305.
30. Talmi-Frank D, Nasereddin A, Schnur LF, Schonian G, Toz SO, Jaffe CL, Baneth G (2010) Detection and identification of old world Leishmania by High Reso-lution Melt Analysis. PLoS Negl Trop Dis. 4(1): e51.
31. Ngui R, Lim YL, Chua KH (2012) Rapid detection and identification of Hook-worm infections through High Reso-lution Melting (HRM) analysis. PloS One. 7(7): e4196.
2. Murray JL, Rosenfeld LC, Lim SS, An-drews KG, Foreman KJ, Haring D, Full-man N, Naghavi M, Lozano R, Lopez AD (2012) Global malaria mortality be-tween 1980 and 2010: A systematic re-view. Lancet. 379: 413–431.
3. De Mast Q, Brouwers J, Syafruddin D, Bousema T, Baidjoe AY, De Groot PG, Van der Ven AJ, Fijnheer R (2015) Is asymptomatic malaria really asympto-mat¬ic? Hematological, vas¬cular and in-flam¬matory effects of asymptomatic malaria parasitaemia. J Infect. 71: 587–596.
4. WHO (2019) World Malaria Report. World Health Organization press. Ge-neva, Swit¬zerland. Available at: https://www.who.int/malaria/en/
5. Nateghpour M, Edrissian GhH, Motevalli Haghi A, Farivar L, Kazemi-Rad E (2015) Diagnosing malaria cases re-ferred to the Malaria Reference La-boratory in Tehran University of Medi-cal Sciences, Iran. Ira¬nian J Para¬sitol. 10: 547–553.
6. Zoghi S, Mehrizi AA, Raeisi A, Hagh-doost AA, Turki H, Safari R, Kahanali AA, Zaker S (2012) Survey of Asymp-tomatic malaria cases in low transmis-sion settings of Iran under elimination pro¬gramme. Malar J. 11: 126–136.
7. Chen I, Sian EC, Gosling R, Hamainza B, Killeen G, Magill A, Omeara W, Price RN, Riley EM (2016) Asymptomatic ma¬laria a chronic and debilitating in-fection that should be treated. Plos Med. 13(1): e1001942.
8. Coleman R, Sattabongkot J, Promstaporm S, Maneechai N, Tippayachai B, Ken-gluecha A, Rachapaew N, Zollner G Scott Miller R, Vaughan J, Thimasarn K, Khun¬tirat B (2006) Comparison of PCR and microscopy for the detection of asymptomatic malaria in a Plasmo-dium falciparum/vivax endemic area in Thailand. Malar J. 5: 121–128.
9. Cooke A, Chiodini PL, Doherty T, Moody AH, Ries J, Pinder M (1999) Compari-son of a parasite lactate dehy¬drogenase-based Immunochromato¬graphic antigen de¬tection assay (opti¬mal) with micros-copy for the detection of malaria para-site in human blood samples. Am J Soc Trop Med Hyg. 60: 173–176.
10. Chua K, Lim S, Ng C, Lee P, Lim YA, Lau, Chai H (2016) Development of High Resolution Melting Analysis for the diagnosis of human malaria. Sci¬en-tific Reports. 5: 15671.
11. Mahende C, Ngasala B, Lusingu J, Yong T, Lushino P, Lemnge M, Mmbando B, Premji Z (2016) Performance of rapid diagnostic test, blood film mi¬croscopy and PCR for diagnosis of malaria infec-tion among febrile chil¬dren from Korog-we district, Tanzania. Malar J. 15: 391–398.
12. Kipanga PN, Omondi D, Mireji PO, Sawa P, Masiga DK, Villinger J (2014) High-resolution analysis reveals low Plasmo-dium parasitaemia infections among mi-cro¬scopically negative fe¬brile patients in western Kenya. Malar J. 13: 429–437.
13. Mangold KA, Manson RU, Koay ES, Ste-phens L, Renger MA, Thomson RB, Pe-terson LR, Kaul KL (2005) Real-time PCR for detection and identifica¬tion of Plasmodium spp. J Clin Micro¬biol. 43: 2435–2440.
14. Canier L, Khim N, Kim S, Sluydts V, Heng S, Dourng D, Eam R, Chy S, Khean C, Loch K, Ken M, Lim H, Siv S, Tho S, Masse-Navette P, Gryseels C, Uk S, Van Roey K, Grietens KP, Sokny M, Thavrin B, Chuor CM, Deubel V, Durnez L, Coosemans M, Menard D (2013) An in¬novative tool for moving malaria PCR detection of parasite reser-voir into the field. Malar J. 12: 405–417.
15. Sargolzaie N, Salehi M, Kiani M, Sakeni M, Hasanzehi A (2011) Malaria epi¬de-miology in Sistan and Baluchestan Prov-ince during April 2008-March 2011, Iran. Zahedan J Res Med Sci. 16: 41–43.
16. Oshaghi MA, Yaghoobi F, Vatandoost H, Abai MR, Akbarzadeh K (2006) Anoph-eles stephensi biological forms, Geo-graph¬ical distribution and malaria transmission in malarious regions of Iran. Pakistan J Biol Sci. 9: 294–298.
17. WHO (2016) Giemsa Staining of Malaria Blood films. Malaria Microscopy Stand-ard Operating Procedure.World Health Organization, Geneva, Swit¬zerland. Avail¬able at: https://apps.who.int/iris/handle/10665/274382
18. Ryan T (2010) Malaria service delivery protocol for sun network. Population services international. Available at: https://silo.tips/download/malaria-ser-vice-delivery-protocol-for-sun-network
19. Bereczky S, Martensson A, Gil JP, Far¬nert A (2005) short report: Rapid DNA ex-traction from archive blood spots on fil-ter paper for genotyping of Plasmo¬dium falciparum. Am J Trop Med Hyg. 72(3): 249–251.
20. Chew CH, Lim YL, Lee PC, Mahmud R, Chua KH (2012) Hexaplex PCR de¬tec-tion system for the identification of five human Plasmodium species with inter-nal controls. J Clin Microbiol. 50(12): 4012–4019.
21. Erali M, Wittwer CT (2010) High Resolu-tion Melting Analysis for gene scan¬ning. PMC. 50: 250–261.
22. Turki H, Raeisi A, Malekzadeh K, Ghan-barnejad A, Zoghi S, Yeryan M, Abedi Nejad M, Mohseni F, Shekari M (2016) Efficiency of Nested-PCR in detecting asymptomatc cases toward malaria elim-ination program in an en¬demic area of Iran. Iran J Parasitol. 10(1): 39–45.
23. Amirshekari MB, Nateghpour M, Raeisi A, Motevalli Haghi A, Farivar L, Edris-sian GhH (2016) Determination of asymp¬tomatic malaria among Af¬ghani and Pakistani immigrants and native population in south of Kerman province, Iran. Iran J Parasitol. 11: 247–252.
24. Nateghpour M, Akbarzadeh K, Farivar L, Amiri A (2011) Detection of asymp¬to-matic malaria among the Afghani immi-grant population in Iranshahr districts of southeastern Iran. Bul Soc Pathol Exot. 104: 321–323.
25. Turki H, Zoghi S, Mehrizi AA, Zakeri S, Raeisi A, Khazan H, Haghdoost AA (2012) Absense of asymptomatic ma-laria infection in endemic area of Ba-shagard District, Hormozgan Prov¬ince, Iran. Ira¬nian J Parasitol. 7: 36–44.
26. Pirahmadi S, Zakeri S, Raeisi A (2017) Ab¬sence of asymptomatic malaria in-fection in a cross-sectional study in Iranshahr District, Iran under elimina-tion pro¬gramme. Iran J Parasitol. 12: 90-100.
27. Gan LS, Loh JP (2010) Rapid identifica-tion of chloroquine and atovaquone drug resistance in Plasmodium falci-parum using High-Resolution Melt Pol-ymerase Chain Reaction. Malar J. 9: 134–139.
28. Hussein EM, Al-Mohammed HI, Hussein AM (2009) Genetic diversity of Dien-tamoeba fragilis isolates of bowel syn-drome patients by High-Resolution Melt¬ing-curve (HRM) analysis. Para¬sitol Res. 105: 1053–1060.
29. Hosseini-Safa AH, Harandi MF, Tajaddini M, Rostami-Nejad M, Mohtashami-Pour M, Pestehchian N (2016) Rapid identi-fication of Echinococcus gran¬ulosus and E. Canadensis using High Resolu-tion Melt¬ing (HRM) analysis by focus-ing on a single nucleotide polymor-phism. J Infect Dis. 69: 300–305.
30. Talmi-Frank D, Nasereddin A, Schnur LF, Schonian G, Toz SO, Jaffe CL, Baneth G (2010) Detection and identification of old world Leishmania by High Reso-lution Melt Analysis. PLoS Negl Trop Dis. 4(1): e51.
31. Ngui R, Lim YL, Chua KH (2012) Rapid detection and identification of Hook-worm infections through High Reso-lution Melting (HRM) analysis. PloS One. 7(7): e4196.
| Files | ||
| Issue | Vol 14 No 4 (2020) | |
| Section | Original Article | |
| DOI | https://doi.org/10.18502/jad.v14i4.5272 | |
| Keywords | ||
| Asymptomatic malaria; High resolution melting (HRM); Microscopy; Rapid diagnostic test (RDTs); Iran | ||
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How to Cite
1.
Ouma FF, Nateghpour M, Motevalli Haghi A, Mohebali M, Farivar L, Hosseini-Safa A, Mosawi SH. Application of High-Resolution Melting (HRM) Technique Towards the Detection of Asymptomatic Malaria in a Malaria Endemic Area of Southeastern Iran Under Elimination Program. J Arthropod Borne Dis. 2021;14(4):353–362.
