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, Rakesh Sehgal, Devendra Bansal, Ali A. Sultan, Ashish Bhalla, Sunit C. Singhi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

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.

Original languageEnglish
Pages (from-to)1374-1381
Number of pages8
JournalAmerican Journal of Tropical Medicine and Hygiene
Volume98
Issue number5
DOIs
Publication statusPublished - 1 Jan 2018

Fingerprint

Vivax Malaria
Plasmodium vivax
Tertiary Care Centers
India
Microscopy
Multiplex Polymerase Chain Reaction
Polymerase Chain Reaction
Early Diagnosis
Sensitivity and Specificity
Indonesia
Gene Targeting
Pakistan
Routine Diagnostic Tests
Limit of Detection
Real-Time Polymerase Chain Reaction
Morbidity
Equipment and Supplies
Mortality

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases
  • Virology

Cite this

Development of visually improved loop mediated isothermal amplification for the diagnosis of plasmodium vivax malaria in a tertiary hospital in Chandigarh, North India. / Kaur, Hargobinder; Sehgal, Rakesh; Bansal, Devendra; Sultan, Ali A.; Bhalla, Ashish; Singhi, Sunit C.

In: American Journal of Tropical Medicine and Hygiene, Vol. 98, No. 5, 01.01.2018, p. 1374-1381.

Research output: Contribution to journalArticle

@article{99b4924abbf447638a5619bfa2baffee,
title = "Development of visually improved loop mediated isothermal amplification for the diagnosis of plasmodium vivax malaria in a tertiary hospital in Chandigarh, North India",
abstract = "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 = "Hargobinder Kaur and Rakesh Sehgal and Devendra Bansal and Sultan, {Ali A.} and Ashish Bhalla and Singhi, {Sunit C.}",
year = "2018",
month = "1",
day = "1",
doi = "10.4269/ajtmh.17-0857",
language = "English",
volume = "98",
pages = "1374--1381",
journal = "American Journal of Tropical Medicine and Hygiene",
issn = "0002-9637",
publisher = "American Society of Tropical Medicine and Hygiene",
number = "5",

}

TY - JOUR

T1 - Development of visually improved loop mediated isothermal amplification for the diagnosis of plasmodium vivax malaria in a tertiary hospital in Chandigarh, North India

AU - Kaur, Hargobinder

AU - Sehgal, Rakesh

AU - Bansal, Devendra

AU - Sultan, Ali A.

AU - Bhalla, Ashish

AU - Singhi, Sunit C.

PY - 2018/1/1

Y1 - 2018/1/1

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=85046905066&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85046905066&partnerID=8YFLogxK

U2 - 10.4269/ajtmh.17-0857

DO - 10.4269/ajtmh.17-0857

M3 - Article

VL - 98

SP - 1374

EP - 1381

JO - American Journal of Tropical Medicine and Hygiene

JF - American Journal of Tropical Medicine and Hygiene

SN - 0002-9637

IS - 5

ER -