Authors:
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- INTRODUCTION
- MATERIAL AND METHODS
- RESULTS
- Subject recruitment and microscopy.
- Demographic and clinical presentation.
- Rapid diagnostic test.
- Comparison of molecular methods with microscopy.
- Visual detection and turnaround time for LAMP assay.
- Estimation of clinical sensitivity and specificity of RDT, multiplex nPCR, RT-PCR, and P. vivax–specific LAMP compared with microscopy.
- Analytical sensitivity and specificity.
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
World Health Organization, 2017. World Malaria Report 2017. Geneva, Switzerland: WHO.
-
2.
Sharma S, Aggarwal KC, Deswal S, Raut D, Roy N, Kapoor R, 2013. The unusual presentation of a usual organism—the changing spectrum of the clinical manifestations of Plasmodium vivax malaria in children: a retrospective study. J Clin Diagn Res 7: 1964–1967.
-
3.
Tjitra E, Anstey NM, Sugiarto P, Warikar N, Kenangalem E, Karyana M, Lampah DA, Price RN, 2008. Multidrug-resistant Plasmodium vivax associated with severe and fatal malaria: a prospective study in Papua, Indonesia. PLoS Med 5: e128.
-
4.
Kochar DK, Das A, Kochar SK, Saxena V, Sirohi P, Garg S, Kochar A, Khatri MP, Gupta V, 2009. Severe Plasmodium vivax malaria: a report on serial cases from Bikaner in northwestern India. Am J Trop Med Hyg 80: 194–198.
-
5.
Fabbri C, de Cássia Mascarenhas-Netto R, Lalwani P, Melo GC, Magalhães BML, Alexandre MAA, Lacerda MVG, Lima ES, 2013. Lipid peroxidation and antioxidant enzymes activity in Plasmodium vivax malaria patients evolving with cholestatic jaundice. Malar J 12: 315.
-
6.
Ketema T, Bacha K, 2013. Plasmodium vivax associated severe malaria complications among children in some malaria endemic areas of Ethiopia. BMC Public Health 13: 637.
-
7.
Kaushik JS, Gomber S, Dewan P, 2012. Clinical and epidemiological profiles of severe malaria in children from Delhi, India. J Health Popul Nutr 30: 113–116.
-
8.
Sharma R, Gohain S, Chandra J, Kumar V, Chopra A, Chatterjee S, Aneja S, Dutta AK, 2012. Plasmodium vivax malaria admissions and risk of mortality in a tertiary-care children’s hospital in north India. Paediatr Int Child Health 32: 152–157.
-
9.
Naha K, Dasari S, Prabhu M, 2012. Spectrum of complications associated with Plasmodium vivax infection in a tertiary hospital in south-western India. Asian Pac J Trop Med 5: 79–82.
-
10.
Chotivanich K, Silamut K, Day NPJ, 2007. Laboratory diagnosis of malaria infection—a short review of methods. NZJ Med Lab Sci 61: 4–7.
-
11.
Lau Y-L, Lai M-Y, Fong M-Y, Jelip J, Mahmud R, 2016. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg 94: 336–339.
-
12.
WHO, 2017. Malaria Rapid Diagnostic Test Performance, Vol. 7. Results of WHO product testing of malaria RDTs: round 7 (2015–2016). Available at: http://www.who.int/malaria/publications/atoz/978924151268/en/.
-
13.
Britton S, Cheng Q, McCarthy JS, 2016. Novel molecular diagnostic tools for malaria elimination: a review of options from the point of view of high-throughput and applicability in resource limited settings. Malar J 15: 88.
-
14.
Perandin F et al. 2004. Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale for routine clinical diagnosis. J Clin Microbiol 42: 1214–1219.
-
15.
Piera KA, Aziz A, William T, Bell D, González IJ, Barber BE, Anstey NM, Grigg MJ, 2017. Detection of Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax using loop-mediated isothermal amplification (LAMP) in a co-endemic area in Malaysia. Malar J 16: 29.
-
16.
Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, do Rosario VE, Thaithong S, Brown KN, 1993. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol 61: 315–320.
-
17.
Hänscheid T, Grobusch MP, 2002. How useful is PCR in the diagnosis of malaria? Trends Parasitol 18: 395–398.
-
18.
Singh R, Singh DP, Savargaonkar D, Singh OP, Bhatt RM, Valecha N, 2017. Evaluation of SYBR green I based visual loop-mediated isothermal amplification (LAMP) assay for genus and species-specific diagnosis of malaria in P. vivax and P. falciparum endemic regions. J Vector Borne Dis 54: 54–60.
-
19.
Patel JC et al. 2014. Field evaluation of a real-time fluorescence loop-mediated isothermal amplification assay, RealAmp, for the diagnosis of malaria in Thailand and India. J Infect Dis 210: 1180–1187.
-
20.
Surabattula R, Vejandla MP, Mallepaddi PC, Faulstich K, Polavarapu R, 2013. Simple, rapid, inexpensive platform for the diagnosis of malaria by loop mediated isothermal amplification (LAMP). Exp Parasitol 134: 333–340.
-
21.
World Health Organization, 2012. Management of Severe Malaria, 3rd edition. Geneva, Switzerland: WHO
-
22.
Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA, 1999. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg 60: 687–692.
-
23.
Han E-T, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 2521–2528.
-
24.
MedCalc, 2015. MedCalc Statistical Software. Version 15.6.1. Ostend, Belgium: MedCalc Software bvba. Available at: https://www.medcalc.org/. Accessed October 5, 2017.
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25.
World Health Organization, 2017. World Malaria Report 2017. Geneva, Switzerland: WHO.
-
26.
Lucchi NW et al. 2016. Evaluation of the illumigene malaria LAMP: a robust molecular diagnostic tool for malaria parasites. Sci Rep 6: 36808.
-
27.
Perera RS, Ding XC, Tully F, Oliver J, Bright N, Bell D, Chiodini PL, Gonzalez IJ, Polley SD, 2017. Development and clinical performance of high throughput loop-mediated isothermal amplification for detection of malaria. PLoS One 12: e0171126.
-
28.
Tao Z-Y et al. 2011. Adaptation of a visualized loop-mediated isothermal amplification technique for field detection of Plasmodium vivax infection. Parasit Vectors 4: 115.
-
29.
Singh R, Savargaonkar D, Bhatt R, Valecha N, 2013. Rapid detection of Plasmodium vivax in saliva and blood using loop mediated isothermal amplification (LAMP) assay. J Infect 67: 245–247.
-
30.
Ghayour Najafabadi Z, Oormazdi H, Akhlaghi L, Meamar AR, Nateghpour M, Farivar L, Razmjou E, 2014. Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: loop-mediated isothermal amplification for malaria diagnosis. Acta Trop 136: 44–49.
-
31.
Valecha N et al. 2009. Histopathology of fatal respiratory distress caused by Plasmodium vivax malaria. Am J Trop Med Hyg 81: 758–762.
-
32.
Mehndiratta S, Rajeshwari K, Dubey AP, 2013. Multiple-organ dysfunction in a case of Plasmodium vivax malaria. J Vector Borne Dis 50: 71–73.
-
33.
Lee H-J, Baek J-H, Chae M-H, Joo H, Lee J-S, Chung M-H, Park Y-K, Kim J-T, 2013. A case of vivax malaria complicated by adult respiratory distress syndrome and successful management with extracorporeal membrane oxygenation. Korean J Parasitol 51: 551–555.
-
34.
Leal-Santos FA, Silva SBR, Crepaldi NP, Nery AF, Martin TOG, Alves-Junior ER, Fontes CJF, 2013. Altered platelet indices as potential markers of severe and complicated malaria caused by Plasmodium vivax: a cross-sectional descriptive study. Malar J 12: 462.
-
35.
Dondorp AM et al. 2005. Estimation of the total parasite biomass in acute falciparum malaria from plasma PfHRP2. PLoS Med 2: e204.
-
36.
Hendriksen ICE et al. 2012. Diagnosing severe falciparum malaria in parasitaemic African children: a prospective evaluation of plasma PfHRP2 measurement. PLoS Med 9: e1001297.
-
37.
Barber BE, William T, Grigg MJ, Parameswaran U, Piera KA, Price RN, Yeo TW, Anstey NM, 2015. Parasite biomass-related inflammation, endothelial activation, microvascular dysfunction and disease severity in vivax malaria. PLoS Pathog 11: e1004558.
-
38.
Ochola LB, Vounatsou P, Smith T, Mabaso ML, Newton CR, 2006. The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard. Lancet Infect Dis 6: 582–588.
-
39.
Murray CK, Gasser RA, Magill AJ, Miller RS, 2008. Update on rapid diagnostic testing for malaria. Clin Microbiol Rev 21: 97–110.
-
40.
Sattabongkot J, Tsuboi T, Han E-T, Bantuchai S, Buates S, 2014. Loop-mediated isothermal amplification assay for rapid diagnosis of malaria infections in an area of endemicity in Thailand. J Clin Microbiol 52: 1471–1477.
-
41.
Singh N, Shukla MM, Shukla MK, Mehra RK, Sharma S, Bharti PK, Singh MP, Singh A, Gunasekar A, 2010. Field and laboratory comparative evaluation of rapid malaria diagnostic tests versus traditional and molecular techniques in India. Malar J 9: 191.
-
42.
Ranjan P, Ghoshal U, 2016. Utility of nested polymerase chain reaction over the microscopy and immuno-chromatographic test in the detection of Plasmodium species and their clinical spectrum. Parasitol Res 115: 3375–3385.
-
43.
Elahi R, Mohon AN, Khan WA, Haque R, Alam MS, 2013. Performance of a HRP-2/pLDH based rapid diagnostic test at the Bangladesh-India-Myanmar border areas for diagnosis of clinical malaria. Malar J 12: 378.
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44.
Kochar DK et al. 2010. Clinical features of children hospitalized with malaria—a study from Bikaner, northwest India. Am J Trop Med Hyg 83: 981–989.
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Poon LLM et al. 2006. Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification. Clin Chem 52: 303–306.
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Development of Visually Improved Loop Mediated Isothermal Amplification for the Diagnosis of Plasmodium vivax Malaria in a Tertiary Hospital in Chandigarh, North India
Hargobinder Kaur
Hargobinder Kaur
Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India;
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Rakesh Sehgal
Rakesh Sehgal
Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India;
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Devendra Bansal
Devendra Bansal
Department of Microbiology and Immunology, Weill Cornell Medicine—Qatar, Cornell University, Qatar Foundation—Education City, Doha, Qatar;
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Ali A. Sultan
Ali A. Sultan
Department of Microbiology and Immunology, Weill Cornell Medicine—Qatar, Cornell University, Qatar Foundation—Education City, Doha, Qatar;
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Ashish Bhalla
Ashish Bhalla
Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India;
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Sunit C. Singhi
Sunit C. Singhi
Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, India
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More than 80% of the global burden of the Plasmodium vivax is contributed by mainly three countries (India, Indonesia, and Pakistan). Reports from last decades have highlighted the occurrence of severe P. vivax malaria which was earlier considered to be benign. The recent trends of increasing P. vivax–associated morbidity and mortality emphasizes the need for early and accurate diagnosis of P. vivax malaria for the timely management of patients. Microscopy is considered a gold standard but needs experienced laboratory technologists. Over the last few years, Polymerase chain reaction (PCR) is being used as a highly sensitive and specific test but it requires expensive equipment which limits its use in the field. Therefore, in the present study, utility of visually improved loop-mediated isothermal amplification (LAMP) for the detection of P. vivax was evaluated targeting 18SrRNA gene in 145 microscopically confirmed P. vivax and 20 P. vivax negative patients. Sensitivity and specificity of LAMP was assessed with respect to microscopy and multiplex nested PCR (nPCR). Results of the LAMP assay was also correlated with rapid diagnostic test, multiplex nPCR and real-time PCR results. Overall, sensitivity and specificity of P. vivax–specific LAMP compared with microscopy were found to be 100% and 85%, respectively. Furthermore, detection limit for LAMP was found to be 0.8 copies/μL and it was also able to detect three complicated cases of P. vivax which were missed by microscopy. This study showed a LAMP assay to be a rapid and very sensitive method for the early diagnosis of both complicated and uncomplicated P. vivax malaria.
Author Notes
Financial support: This publication was made possible by partial financial assistance from NPRP grant (NPRP 5-098-3-021) from the Qatar National Research Fund (a member of Qatar Foundation).
Authors’ addresses: Hargobinder Kaur and Rakesh Sehgal, Department of Medical Parasitology, Postgraduate Institute of Medical Education and Research, Chandigarh, India, E-mails: hargobinderkaur27785@gmail.com and sehgalpgi@gmail.com. Devendra Bansal and Ali A. Sultan, Department of Microbiology and Immunology, Weill Cornell Medicine—Qatar, Cornell University, Qatar Foundation—Education City, Doha, Qatar, E-mails: dbansal2006@gmail.com and als2026@qatar-med.cornell.edu. Ashish Bhalla, Department of Internal Medicine, Postgraduate Institute of Medical Education and Research, Chandigarh, India, E-mail: bhalla.chd@gmail.com. Sunit C. Singhi, Department of Pediatrics, Postgraduate Institute of Medical Education and Research, Chandigarh, India, E-mail: sunit.singhi@gmail.com.
- INTRODUCTION
- MATERIAL AND METHODS
- RESULTS
- Subject recruitment and microscopy.
- Demographic and clinical presentation.
- Rapid diagnostic test.
- Comparison of molecular methods with microscopy.
- Visual detection and turnaround time for LAMP assay.
- Estimation of clinical sensitivity and specificity of RDT, multiplex nPCR, RT-PCR, and P. vivax–specific LAMP compared with microscopy.
- Analytical sensitivity and specificity.
- DISCUSSION
- CONCLUSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1.
World Health Organization, 2017. World Malaria Report 2017. Geneva, Switzerland: WHO.
-
2.
Sharma S, Aggarwal KC, Deswal S, Raut D, Roy N, Kapoor R, 2013. The unusual presentation of a usual organism—the changing spectrum of the clinical manifestations of Plasmodium vivax malaria in children: a retrospective study. J Clin Diagn Res 7: 1964–1967.
-
3.
Tjitra E, Anstey NM, Sugiarto P, Warikar N, Kenangalem E, Karyana M, Lampah DA, Price RN, 2008. Multidrug-resistant Plasmodium vivax associated with severe and fatal malaria: a prospective study in Papua, Indonesia. PLoS Med 5: e128.
-
4.
Kochar DK, Das A, Kochar SK, Saxena V, Sirohi P, Garg S, Kochar A, Khatri MP, Gupta V, 2009. Severe Plasmodium vivax malaria: a report on serial cases from Bikaner in northwestern India. Am J Trop Med Hyg 80: 194–198.
-
5.
Fabbri C, de Cássia Mascarenhas-Netto R, Lalwani P, Melo GC, Magalhães BML, Alexandre MAA, Lacerda MVG, Lima ES, 2013. Lipid peroxidation and antioxidant enzymes activity in Plasmodium vivax malaria patients evolving with cholestatic jaundice. Malar J 12: 315.
-
6.
Ketema T, Bacha K, 2013. Plasmodium vivax associated severe malaria complications among children in some malaria endemic areas of Ethiopia. BMC Public Health 13: 637.
-
7.
Kaushik JS, Gomber S, Dewan P, 2012. Clinical and epidemiological profiles of severe malaria in children from Delhi, India. J Health Popul Nutr 30: 113–116.
-
8.
Sharma R, Gohain S, Chandra J, Kumar V, Chopra A, Chatterjee S, Aneja S, Dutta AK, 2012. Plasmodium vivax malaria admissions and risk of mortality in a tertiary-care children’s hospital in north India. Paediatr Int Child Health 32: 152–157.
-
9.
Naha K, Dasari S, Prabhu M, 2012. Spectrum of complications associated with Plasmodium vivax infection in a tertiary hospital in south-western India. Asian Pac J Trop Med 5: 79–82.
-
10.
Chotivanich K, Silamut K, Day NPJ, 2007. Laboratory diagnosis of malaria infection—a short review of methods. NZJ Med Lab Sci 61: 4–7.
-
11.
Lau Y-L, Lai M-Y, Fong M-Y, Jelip J, Mahmud R, 2016. Loop-mediated isothermal amplification assay for identification of five human Plasmodium species in Malaysia. Am J Trop Med Hyg 94: 336–339.
-
12.
WHO, 2017. Malaria Rapid Diagnostic Test Performance, Vol. 7. Results of WHO product testing of malaria RDTs: round 7 (2015–2016). Available at: http://www.who.int/malaria/publications/atoz/978924151268/en/.
-
13.
Britton S, Cheng Q, McCarthy JS, 2016. Novel molecular diagnostic tools for malaria elimination: a review of options from the point of view of high-throughput and applicability in resource limited settings. Malar J 15: 88.
-
14.
Perandin F et al. 2004. Development of a real-time PCR assay for detection of Plasmodium falciparum, Plasmodium vivax, and Plasmodium ovale for routine clinical diagnosis. J Clin Microbiol 42: 1214–1219.
-
15.
Piera KA, Aziz A, William T, Bell D, González IJ, Barber BE, Anstey NM, Grigg MJ, 2017. Detection of Plasmodium knowlesi, Plasmodium falciparum and Plasmodium vivax using loop-mediated isothermal amplification (LAMP) in a co-endemic area in Malaysia. Malar J 16: 29.
-
16.
Snounou G, Viriyakosol S, Zhu XP, Jarra W, Pinheiro L, do Rosario VE, Thaithong S, Brown KN, 1993. High sensitivity of detection of human malaria parasites by the use of nested polymerase chain reaction. Mol Biochem Parasitol 61: 315–320.
-
17.
Hänscheid T, Grobusch MP, 2002. How useful is PCR in the diagnosis of malaria? Trends Parasitol 18: 395–398.
-
18.
Singh R, Singh DP, Savargaonkar D, Singh OP, Bhatt RM, Valecha N, 2017. Evaluation of SYBR green I based visual loop-mediated isothermal amplification (LAMP) assay for genus and species-specific diagnosis of malaria in P. vivax and P. falciparum endemic regions. J Vector Borne Dis 54: 54–60.
-
19.
Patel JC et al. 2014. Field evaluation of a real-time fluorescence loop-mediated isothermal amplification assay, RealAmp, for the diagnosis of malaria in Thailand and India. J Infect Dis 210: 1180–1187.
-
20.
Surabattula R, Vejandla MP, Mallepaddi PC, Faulstich K, Polavarapu R, 2013. Simple, rapid, inexpensive platform for the diagnosis of malaria by loop mediated isothermal amplification (LAMP). Exp Parasitol 134: 333–340.
-
21.
World Health Organization, 2012. Management of Severe Malaria, 3rd edition. Geneva, Switzerland: WHO
-
22.
Singh B, Bobogare A, Cox-Singh J, Snounou G, Abdullah MS, Rahman HA, 1999. A genus- and species-specific nested polymerase chain reaction malaria detection assay for epidemiologic studies. Am J Trop Med Hyg 60: 687–692.
-
23.
Han E-T, Watanabe R, Sattabongkot J, Khuntirat B, Sirichaisinthop J, Iriko H, Jin L, Takeo S, Tsuboi T, 2007. Detection of four Plasmodium species by genus- and species-specific loop-mediated isothermal amplification for clinical diagnosis. J Clin Microbiol 45: 2521–2528.
-
24.
MedCalc, 2015. MedCalc Statistical Software. Version 15.6.1. Ostend, Belgium: MedCalc Software bvba. Available at: https://www.medcalc.org/. Accessed October 5, 2017.
-
25.
World Health Organization, 2017. World Malaria Report 2017. Geneva, Switzerland: WHO.
-
26.
Lucchi NW et al. 2016. Evaluation of the illumigene malaria LAMP: a robust molecular diagnostic tool for malaria parasites. Sci Rep 6: 36808.
-
27.
Perera RS, Ding XC, Tully F, Oliver J, Bright N, Bell D, Chiodini PL, Gonzalez IJ, Polley SD, 2017. Development and clinical performance of high throughput loop-mediated isothermal amplification for detection of malaria. PLoS One 12: e0171126.
-
28.
Tao Z-Y et al. 2011. Adaptation of a visualized loop-mediated isothermal amplification technique for field detection of Plasmodium vivax infection. Parasit Vectors 4: 115.
-
29.
Singh R, Savargaonkar D, Bhatt R, Valecha N, 2013. Rapid detection of Plasmodium vivax in saliva and blood using loop mediated isothermal amplification (LAMP) assay. J Infect 67: 245–247.
-
30.
Ghayour Najafabadi Z, Oormazdi H, Akhlaghi L, Meamar AR, Nateghpour M, Farivar L, Razmjou E, 2014. Detection of Plasmodium vivax and Plasmodium falciparum DNA in human saliva and urine: loop-mediated isothermal amplification for malaria diagnosis. Acta Trop 136: 44–49.
-
31.
Valecha N et al. 2009. Histopathology of fatal respiratory distress caused by Plasmodium vivax malaria. Am J Trop Med Hyg 81: 758–762.
-
32.
Mehndiratta S, Rajeshwari K, Dubey AP, 2013. Multiple-organ dysfunction in a case of Plasmodium vivax malaria. J Vector Borne Dis 50: 71–73.
-
33.
Lee H-J, Baek J-H, Chae M-H, Joo H, Lee J-S, Chung M-H, Park Y-K, Kim J-T, 2013. A case of vivax malaria complicated by adult respiratory distress syndrome and successful management with extracorporeal membrane oxygenation. Korean J Parasitol 51: 551–555.
-
34.
Leal-Santos FA, Silva SBR, Crepaldi NP, Nery AF, Martin TOG, Alves-Junior ER, Fontes CJF, 2013. Altered platelet indices as potential markers of severe and complicated malaria caused by Plasmodium vivax: a cross-sectional descriptive study. Malar J 12: 462.
-
35.
Dondorp AM et al. 2005. Estimation of the total parasite biomass in acute falciparum malaria from plasma PfHRP2. PLoS Med 2: e204.
-
36.
Hendriksen ICE et al. 2012. Diagnosing severe falciparum malaria in parasitaemic African children: a prospective evaluation of plasma PfHRP2 measurement. PLoS Med 9: e1001297.
-
37.
Barber BE, William T, Grigg MJ, Parameswaran U, Piera KA, Price RN, Yeo TW, Anstey NM, 2015. Parasite biomass-related inflammation, endothelial activation, microvascular dysfunction and disease severity in vivax malaria. PLoS Pathog 11: e1004558.
-
38.
Ochola LB, Vounatsou P, Smith T, Mabaso ML, Newton CR, 2006. The reliability of diagnostic techniques in the diagnosis and management of malaria in the absence of a gold standard. Lancet Infect Dis 6: 582–588.
-
39.
Murray CK, Gasser RA, Magill AJ, Miller RS, 2008. Update on rapid diagnostic testing for malaria. Clin Microbiol Rev 21: 97–110.
-
40.
Sattabongkot J, Tsuboi T, Han E-T, Bantuchai S, Buates S, 2014. Loop-mediated isothermal amplification assay for rapid diagnosis of malaria infections in an area of endemicity in Thailand. J Clin Microbiol 52: 1471–1477.
-
41.
Singh N, Shukla MM, Shukla MK, Mehra RK, Sharma S, Bharti PK, Singh MP, Singh A, Gunasekar A, 2010. Field and laboratory comparative evaluation of rapid malaria diagnostic tests versus traditional and molecular techniques in India. Malar J 9: 191.
-
42.
Ranjan P, Ghoshal U, 2016. Utility of nested polymerase chain reaction over the microscopy and immuno-chromatographic test in the detection of Plasmodium species and their clinical spectrum. Parasitol Res 115: 3375–3385.
-
43.
Elahi R, Mohon AN, Khan WA, Haque R, Alam MS, 2013. Performance of a HRP-2/pLDH based rapid diagnostic test at the Bangladesh-India-Myanmar border areas for diagnosis of clinical malaria. Malar J 12: 378.
-
44.
Kochar DK et al. 2010. Clinical features of children hospitalized with malaria—a study from Bikaner, northwest India. Am J Trop Med Hyg 83: 981–989.
-
45.
Poon LLM et al. 2006. Sensitive and inexpensive molecular test for falciparum malaria: detecting Plasmodium falciparum DNA directly from heat-treated blood by loop-mediated isothermal amplification. Clin Chem 52: 303–306.
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