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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.

Abstract Loop-mediated isothermal amplification (LAMP) is an emerging nucleic acid based diag- nostic approach that is easily adaptable to the field settings with limited technical resources. This study was aimed to evaluate the LAMP assay for the detection and identification of Plasmodium falciparum and P. vivax infection in malaria suspected cases using genus and species-specific assay.
PMID
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Authors

Mayor MeshTerms
Keywords
Journal Title journal of vector borne diseases
Publication Year Start




PMID- 28352046
OWN - NLM
STAT- MEDLINE
DA  - 20170329
DCOM- 20170414
LR  - 20170414
IS  - 0972-9062 (Print)
IS  - 0972-9062 (Linking)
VI  - 54
IP  - 1
DP  - 2017 Jan-Mar
TI  - 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.
PG  - 54-60
AB  - BACKGROUND & OBJECTIVES: Loop-mediated isothermal amplification (LAMP) is an
      emerging nucleic acid based diag- nostic approach that is easily adaptable to the
      field settings with limited technical resources. This study was aimed to evaluate
      the LAMP assay for the detection and identification of Plasmodium falciparum and 
      P. vivax infection in malaria suspected cases using genus and species-specific
      assay. METHODS: The 18S rRNA-based LAMP assay was evaluated for diagnosis of
      genus Plasmodium, and species- specific diagnosis of P. falciparum and P. vivax, 
      infection employing 317 malaria suspected cases, and the results were compared
      with those obtained by 18S nested PCR (n-PCR). All the samples were confirmed by 
      microscopy for the presence of Plasmodium parasite. RESULTS: The n-PCR was
      positive in all Plasmodium-infected cases (n=257; P. falciparum=133; P.
      vivax=124) and negative in microscopy negative cases (n=58) except for two cases 
      which were positive for P. vivax, giving a sen- sitivity of 100% (95% CI:
      97.04-100%) and a specificity of 100% (95% CI: 88.45-99.5%). Genus-specific LAMP 
      assay missed 11 (3.2%) microscopy and n-PCR confirmed vivax malaria cases.
      Considering PCR results as a refer- ence, LAMP was 100% sensitive and specific
      for P. falciparum, whereas it exhibited 95.16% sensitivity and 96.7% specificity 
      for P. vivax. The n-PCR assay detected 10 mixed infection cases while
      species-specific LAMP detected five mixed infection cases of P. vivax and P.
      falciparum, which were not detected by microscopy. INTERPRETATION & CONCLUSION:
      Genus-specific LAMP assay displayed low sensitivity. Falciparum specific LAMP
      assay displayed high sensitivity whereas vivax specific LAMP assay displayed low 
      sensitivity. Failed detection of vivax cases otherwise confirmed by the n-PCR
      assay indicates exploitation of new targets and improved detection methods to
      attain 100% results for P. vivax detection.
FAU - Singh, Ruchi
AU  - Singh R
AD  - National Institute of Malaria Research (ICMR), Dwarka; National Institute of
      Pathology (ICMR), Safdarjung Hospital Campus, New Delhi, India.
FAU - Singh, Dhirendra Pratap
AU  - Singh DP
AD  - National Institute of Malaria Research (ICMR), Dwarka, India.
FAU - Savargaonkar, Deepali
AU  - Savargaonkar D
AD  - National Institute of Malaria Research (ICMR), Dwarka, India.
FAU - Singh, Om P
AU  - Singh OP
AD  - National Institute of Malaria Research (ICMR), Dwarka, India.
FAU - Bhatt, Rajendra M
AU  - Bhatt RM
AD  - National Institute of Malaria Research, Field Unit, Raipur, Chhattisgarh, India.
FAU - Valecha, Neena
AU  - Valecha N
AD  - National Institute of Malaria Research (ICMR), Dwarka, India.
LA  - eng
PT  - Comparative Study
PT  - Evaluation Studies
PT  - Journal Article
PL  - India
TA  - J Vector Borne Dis
JT  - Journal of vector borne diseases
JID - 101212761
RN  - 0 (Organic Chemicals)
RN  - 0 (RNA, Ribosomal, 18S)
RN  - 163795-75-3 (SYBR Green I)
SB  - IM
MH  - Adolescent
MH  - Adult
MH  - Aged
MH  - Aged, 80 and over
MH  - Child
MH  - Child, Preschool
MH  - Female
MH  - Humans
MH  - Infant
MH  - Malaria, Falciparum/*diagnosis/parasitology
MH  - Malaria, Vivax/*diagnosis/parasitology
MH  - Male
MH  - Middle Aged
MH  - Molecular Diagnostic Techniques/*methods
MH  - Nucleic Acid Amplification Techniques/*methods
MH  - Organic Chemicals/analysis
MH  - Plasmodium falciparum/genetics/*isolation & purification
MH  - Plasmodium vivax/genetics/*isolation & purification
MH  - RNA, Ribosomal, 18S/genetics
MH  - Sensitivity and Specificity
MH  - Staining and Labeling/*methods
MH  - Young Adult
EDAT- 2017/03/30 06:00
MHDA- 2017/04/15 06:00
CRDT- 2017/03/30 06:00
AID - JVectorBorneDis_2017_54_1_54_203184 [pii]
PST - ppublish
SO  - J Vector Borne Dis. 2017 Jan-Mar;54(1):54-60.

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