Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India

IntroductionThe diagnosis of infectious reservoirs in malaria (gametocytes) is necessary, especially in low-density infections and asymptomatic malaria patients. The gametocyte stage is a surrogate marker for infection of P. falciparum malaria in healthy individuals. The early detection of infectiou...

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Main Authors: Ram Das, Kapil Vashisht, Lokesh Kori, Kuldeep Singh, Gaurav Kumar, Izazul Hasan, Jugal Gam, Kailash C. Pandey
Format: Article
Language:English
Published: Frontiers Media S.A. 2024-03-01
Series:Frontiers in Tropical Diseases
Subjects:
Online Access:https://www.frontiersin.org/articles/10.3389/fitd.2024.1366462/full
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author Ram Das
Kapil Vashisht
Lokesh Kori
Kuldeep Singh
Gaurav Kumar
Izazul Hasan
Jugal Gam
Kailash C. Pandey
author_facet Ram Das
Kapil Vashisht
Lokesh Kori
Kuldeep Singh
Gaurav Kumar
Izazul Hasan
Jugal Gam
Kailash C. Pandey
author_sort Ram Das
collection DOAJ
description IntroductionThe diagnosis of infectious reservoirs in malaria (gametocytes) is necessary, especially in low-density infections and asymptomatic malaria patients. The gametocyte stage is a surrogate marker for infection of P. falciparum malaria in healthy individuals. The early detection of infectious gametocytes and treatment will strengthen our efforts in curbing transmission. The nested PCR and real-time quantitative PCR (RT-qPCR) methods have been demonstrated for the diagnosis of infectious gametocyte reservoirs. In this study, RDT, blood smear microscopy, and nested-PCR were used for the detection of P. falciparum and P. vivax, and compared with RT-qPCR detection of Pfg27 gametocyte biomarker gene.MethodsIn the present cross-sectional study, 356 human blood samples were collected from endemic areas of Kokrajhar Assam (asymptomatic and symptomatic malaria patients) for malaria diagnosis.ResultsA total of 8.42%(30/356) incidence of malaria was observed. Malaria patients were observed to be both symptomatic, 80%(24/30; 13Pf+11Pv), and asymptomatic, 20%(6 (4Pf +2Pv)). More than 64%(11/17) of Pf and 92.3%(12/13) of Pv infections were observed in children and the adolescent population (age <20 years) by RDT, microscopy, nested PCR, and RT-qPCR methods. The prevalence of Pf infection was 4.77%(17/356) by RT-qPCR method. Of 16 the Pf positive samples 81.25%(13/16) were symptomatic and 18.75%(3/16) were asymptomatic. One asymptomatic individual was found positive for Pf infection by the RT-qPCR method.ConclusionThe findings from this research study revealed that the routine microscopy and RDT methods are insufficient for detecting all asymptomatic malaria and gametocyte infectious reservoirs. The early detection of infectious P. falciparum gametocytes and the treatment of patients will be helpful in preventing the transmission of malaria.
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spelling doaj.art-7ca75ed38f40445e90e3117a4edc37fa2024-03-07T09:12:19ZengFrontiers Media S.A.Frontiers in Tropical Diseases2673-75152024-03-01510.3389/fitd.2024.13664621366462Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern IndiaRam Das0Kapil Vashisht1Lokesh Kori2Kuldeep Singh3Gaurav Kumar4Izazul Hasan5Jugal Gam6Kailash C. Pandey7Parasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaHealth Services, District Malaria Office Kokrajhar, Assam, IndiaParasite Host Biology Group, ICMR-National Institute of Malaria Research, Delhi, IndiaIntroductionThe diagnosis of infectious reservoirs in malaria (gametocytes) is necessary, especially in low-density infections and asymptomatic malaria patients. The gametocyte stage is a surrogate marker for infection of P. falciparum malaria in healthy individuals. The early detection of infectious gametocytes and treatment will strengthen our efforts in curbing transmission. The nested PCR and real-time quantitative PCR (RT-qPCR) methods have been demonstrated for the diagnosis of infectious gametocyte reservoirs. In this study, RDT, blood smear microscopy, and nested-PCR were used for the detection of P. falciparum and P. vivax, and compared with RT-qPCR detection of Pfg27 gametocyte biomarker gene.MethodsIn the present cross-sectional study, 356 human blood samples were collected from endemic areas of Kokrajhar Assam (asymptomatic and symptomatic malaria patients) for malaria diagnosis.ResultsA total of 8.42%(30/356) incidence of malaria was observed. Malaria patients were observed to be both symptomatic, 80%(24/30; 13Pf+11Pv), and asymptomatic, 20%(6 (4Pf +2Pv)). More than 64%(11/17) of Pf and 92.3%(12/13) of Pv infections were observed in children and the adolescent population (age <20 years) by RDT, microscopy, nested PCR, and RT-qPCR methods. The prevalence of Pf infection was 4.77%(17/356) by RT-qPCR method. Of 16 the Pf positive samples 81.25%(13/16) were symptomatic and 18.75%(3/16) were asymptomatic. One asymptomatic individual was found positive for Pf infection by the RT-qPCR method.ConclusionThe findings from this research study revealed that the routine microscopy and RDT methods are insufficient for detecting all asymptomatic malaria and gametocyte infectious reservoirs. The early detection of infectious P. falciparum gametocytes and the treatment of patients will be helpful in preventing the transmission of malaria.https://www.frontiersin.org/articles/10.3389/fitd.2024.1366462/fullPlasmodium falciparummalariaRT-qPCRgametocyte detectionPfg27- gametocyte biomarker gene
spellingShingle Ram Das
Kapil Vashisht
Lokesh Kori
Kuldeep Singh
Gaurav Kumar
Izazul Hasan
Jugal Gam
Kailash C. Pandey
Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
Frontiers in Tropical Diseases
Plasmodium falciparum
malaria
RT-qPCR
gametocyte detection
Pfg27- gametocyte biomarker gene
title Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
title_full Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
title_fullStr Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
title_full_unstemmed Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
title_short Detection of the infective Plasmodium falciparum gametocytes by RT-qPCR assay from a malaria-endemic region of Northeastern India
title_sort detection of the infective plasmodium falciparum gametocytes by rt qpcr assay from a malaria endemic region of northeastern india
topic Plasmodium falciparum
malaria
RT-qPCR
gametocyte detection
Pfg27- gametocyte biomarker gene
url https://www.frontiersin.org/articles/10.3389/fitd.2024.1366462/full
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