Point of care diagnosis of multiple schistosome parasites: Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP)

Nilanjan Lodh, Kei Mikita, Kwabena M. Bosompem, William K. Anyan, Joseph K. Quartey, Joseph Otchere, Clive J. Shiff

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness. While serology, such as antigen capture tests might improve sensitivity, it has been shown that the presence of species-specific DNA fragments will indicate, most effectively, the presence of active parasites. Polymerase chain reaction (PCR) can amplify and detect DNA from urine residue captured on Whatman No. 3 filter paper that is dried after filtration. Previously we have detected S. mansoni and S. haematobium parasite-specific small repeat DNA fragment from filtered urine on filter paper by PCR. In the current study, we assessed the efficacy of detection of 86 urine samples for either or both schistosome parasites by PCR and loop-mediated isothermal amplification (LAMP) that were collected from a low to moderate transmission area in Ghana. Two different DNA extraction methods, standard extraction kit and field usable LAMP-PURE kit were also evaluated by PCR and LAMP amplification. With S. haematobium LAMP amplification for both extractions showed similar sensitivity and specificity when compared with PCR amplification (100%) verified by gel electrophoresis. For S. mansoni sensitivity was highest for LAMP amplification (100%) for standard extraction than PCR and LAMP with LAMP-PURE (99% and 94%). The LAMP-PURE extraction produced false negatives, which require further investigation for this field usable extraction kit. Overall high positive and negative predictive values (90% − 100%) for both species demonstrated a highly robust approach. The LAMP approach is close to point of care use and equally sensitive and specific to detection of species-specific DNA by PCR. LAMP can be an effective means to detect low intensity infection due to its simplicity and minimal DNA extraction requirement. This will enhance the effectiveness of surveillance and MDA control programs of schistosomiasis.

Original languageEnglish
Pages (from-to)125-129
Number of pages5
JournalActa Tropica
Volume173
DOIs
Publication statusPublished - 2017 Sep 1

Fingerprint

Point-of-Care Systems
Schistosoma
Parasites
urine
Urine
parasites
Polymerase Chain Reaction
DNA
polymerase chain reaction
Haematobia
Praziquantel
Ghana
Food Chain
Drug and Narcotic Control
Schistosomiasis
Snails
Program Evaluation
Serology
DNA-Directed DNA Polymerase
Infection Control

Keywords

  • LAMP
  • LAMP-PURE
  • PCR
  • Repeat DNA
  • Schistosomiasis
  • Urine

ASJC Scopus subject areas

  • Parasitology
  • Infectious Diseases

Cite this

Point of care diagnosis of multiple schistosome parasites : Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP). / Lodh, Nilanjan; Mikita, Kei; Bosompem, Kwabena M.; Anyan, William K.; Quartey, Joseph K.; Otchere, Joseph; Shiff, Clive J.

In: Acta Tropica, Vol. 173, 01.09.2017, p. 125-129.

Research output: Contribution to journalArticle

Lodh, Nilanjan ; Mikita, Kei ; Bosompem, Kwabena M. ; Anyan, William K. ; Quartey, Joseph K. ; Otchere, Joseph ; Shiff, Clive J. / Point of care diagnosis of multiple schistosome parasites : Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP). In: Acta Tropica. 2017 ; Vol. 173. pp. 125-129.
@article{8499786e164d4833844aa55a2b015a4b,
title = "Point of care diagnosis of multiple schistosome parasites: Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP)",
abstract = "Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness. While serology, such as antigen capture tests might improve sensitivity, it has been shown that the presence of species-specific DNA fragments will indicate, most effectively, the presence of active parasites. Polymerase chain reaction (PCR) can amplify and detect DNA from urine residue captured on Whatman No. 3 filter paper that is dried after filtration. Previously we have detected S. mansoni and S. haematobium parasite-specific small repeat DNA fragment from filtered urine on filter paper by PCR. In the current study, we assessed the efficacy of detection of 86 urine samples for either or both schistosome parasites by PCR and loop-mediated isothermal amplification (LAMP) that were collected from a low to moderate transmission area in Ghana. Two different DNA extraction methods, standard extraction kit and field usable LAMP-PURE kit were also evaluated by PCR and LAMP amplification. With S. haematobium LAMP amplification for both extractions showed similar sensitivity and specificity when compared with PCR amplification (100{\%}) verified by gel electrophoresis. For S. mansoni sensitivity was highest for LAMP amplification (100{\%}) for standard extraction than PCR and LAMP with LAMP-PURE (99{\%} and 94{\%}). The LAMP-PURE extraction produced false negatives, which require further investigation for this field usable extraction kit. Overall high positive and negative predictive values (90{\%} − 100{\%}) for both species demonstrated a highly robust approach. The LAMP approach is close to point of care use and equally sensitive and specific to detection of species-specific DNA by PCR. LAMP can be an effective means to detect low intensity infection due to its simplicity and minimal DNA extraction requirement. This will enhance the effectiveness of surveillance and MDA control programs of schistosomiasis.",
keywords = "LAMP, LAMP-PURE, PCR, Repeat DNA, Schistosomiasis, Urine",
author = "Nilanjan Lodh and Kei Mikita and Bosompem, {Kwabena M.} and Anyan, {William K.} and Quartey, {Joseph K.} and Joseph Otchere and Shiff, {Clive J.}",
year = "2017",
month = "9",
day = "1",
doi = "10.1016/j.actatropica.2017.06.015",
language = "English",
volume = "173",
pages = "125--129",
journal = "Acta Tropica",
issn = "0001-706X",
publisher = "Elsevier",

}

TY - JOUR

T1 - Point of care diagnosis of multiple schistosome parasites

T2 - Species-specific DNA detection in urine by loop-mediated isothermal amplification (LAMP)

AU - Lodh, Nilanjan

AU - Mikita, Kei

AU - Bosompem, Kwabena M.

AU - Anyan, William K.

AU - Quartey, Joseph K.

AU - Otchere, Joseph

AU - Shiff, Clive J.

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness. While serology, such as antigen capture tests might improve sensitivity, it has been shown that the presence of species-specific DNA fragments will indicate, most effectively, the presence of active parasites. Polymerase chain reaction (PCR) can amplify and detect DNA from urine residue captured on Whatman No. 3 filter paper that is dried after filtration. Previously we have detected S. mansoni and S. haematobium parasite-specific small repeat DNA fragment from filtered urine on filter paper by PCR. In the current study, we assessed the efficacy of detection of 86 urine samples for either or both schistosome parasites by PCR and loop-mediated isothermal amplification (LAMP) that were collected from a low to moderate transmission area in Ghana. Two different DNA extraction methods, standard extraction kit and field usable LAMP-PURE kit were also evaluated by PCR and LAMP amplification. With S. haematobium LAMP amplification for both extractions showed similar sensitivity and specificity when compared with PCR amplification (100%) verified by gel electrophoresis. For S. mansoni sensitivity was highest for LAMP amplification (100%) for standard extraction than PCR and LAMP with LAMP-PURE (99% and 94%). The LAMP-PURE extraction produced false negatives, which require further investigation for this field usable extraction kit. Overall high positive and negative predictive values (90% − 100%) for both species demonstrated a highly robust approach. The LAMP approach is close to point of care use and equally sensitive and specific to detection of species-specific DNA by PCR. LAMP can be an effective means to detect low intensity infection due to its simplicity and minimal DNA extraction requirement. This will enhance the effectiveness of surveillance and MDA control programs of schistosomiasis.

AB - Schistosomes are easily transmitted and high chance of repeat infection, so if control strategies based on targeted mass drug administration (MDA) are to succeed it is essential to have a test that is sensitive, accurate and simple to use. It is known and regularly demonstrated that praziquantel does not always eliminate an infection so in spite of the successes of control programs a residual of the reservoir survives to re-infect snails. The issue of diagnostic sensitivity becomes more critical in the assessment of program effectiveness. While serology, such as antigen capture tests might improve sensitivity, it has been shown that the presence of species-specific DNA fragments will indicate, most effectively, the presence of active parasites. Polymerase chain reaction (PCR) can amplify and detect DNA from urine residue captured on Whatman No. 3 filter paper that is dried after filtration. Previously we have detected S. mansoni and S. haematobium parasite-specific small repeat DNA fragment from filtered urine on filter paper by PCR. In the current study, we assessed the efficacy of detection of 86 urine samples for either or both schistosome parasites by PCR and loop-mediated isothermal amplification (LAMP) that were collected from a low to moderate transmission area in Ghana. Two different DNA extraction methods, standard extraction kit and field usable LAMP-PURE kit were also evaluated by PCR and LAMP amplification. With S. haematobium LAMP amplification for both extractions showed similar sensitivity and specificity when compared with PCR amplification (100%) verified by gel electrophoresis. For S. mansoni sensitivity was highest for LAMP amplification (100%) for standard extraction than PCR and LAMP with LAMP-PURE (99% and 94%). The LAMP-PURE extraction produced false negatives, which require further investigation for this field usable extraction kit. Overall high positive and negative predictive values (90% − 100%) for both species demonstrated a highly robust approach. The LAMP approach is close to point of care use and equally sensitive and specific to detection of species-specific DNA by PCR. LAMP can be an effective means to detect low intensity infection due to its simplicity and minimal DNA extraction requirement. This will enhance the effectiveness of surveillance and MDA control programs of schistosomiasis.

KW - LAMP

KW - LAMP-PURE

KW - PCR

KW - Repeat DNA

KW - Schistosomiasis

KW - Urine

UR - http://www.scopus.com/inward/record.url?scp=85020525412&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85020525412&partnerID=8YFLogxK

U2 - 10.1016/j.actatropica.2017.06.015

DO - 10.1016/j.actatropica.2017.06.015

M3 - Article

C2 - 28619672

AN - SCOPUS:85020525412

VL - 173

SP - 125

EP - 129

JO - Acta Tropica

JF - Acta Tropica

SN - 0001-706X

ER -