Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device
LAMP diagnosis of malaria is simple and cost-effective with acceptable sensitivity and specificity as compared to standard diagnostic modules such as microscopy, RDTs and nested PCR, and thus its deployment for onsite screening of malaria in resource-limited regions is under consideration. However,...
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Frontiers Media S.A.
2022-08-01
|
| Series: | Frontiers in Cellular and Infection Microbiology |
| Subjects: | |
| Online Access: | https://www.frontiersin.org/articles/10.3389/fcimb.2022.961832/full |
| _version_ | 1811339775587647488 |
|---|---|
| author | Madhu Puri Harsimran Kaur Brar Evanka Madan Rajesh Srinivasan Kapil Rawat Sai Siva Gorthi Geeta Kumari Raj Sah Sashi Bhusan Ojha Subhendu Panigrahi Gunanidhi Dhangadamajhi Rohini Muthuswami Shailja Singh Rentala Madhubala |
| author_facet | Madhu Puri Harsimran Kaur Brar Evanka Madan Rajesh Srinivasan Kapil Rawat Sai Siva Gorthi Geeta Kumari Raj Sah Sashi Bhusan Ojha Subhendu Panigrahi Gunanidhi Dhangadamajhi Rohini Muthuswami Shailja Singh Rentala Madhubala |
| author_sort | Madhu Puri |
| collection | DOAJ |
| description | LAMP diagnosis of malaria is simple and cost-effective with acceptable sensitivity and specificity as compared to standard diagnostic modules such as microscopy, RDTs and nested PCR, and thus its deployment for onsite screening of malaria in resource-limited regions is under consideration. However, the requirement of an electricity-operated dry bath and bulky read-out unit is still a major concern. In an effort to simplify this limitation, we have developed a portable LAMP device and fluorescence readout unit which can be used in the rapid point-of-care diagnosis of malaria. We have developed a point-of-care diagnostic LAMP device that is easy to operate by a mobile application, and the results can be quantified with a fluorescent readout unit. The diagnostic performance of the device was evaluated in 90 P. falciparum-infected clinical isolates stored at 4°C for 6-7 years and 10 freshly collected isolates from healthy volunteers. The LOD and quantitative ability of LAMP in estimating parasitemia levels were revealed with laboratory-grown P. falciparum strain (3D7). The LAMP assay performed in our device was exclusive for P. falciparum detection with sensitivity and specificity determined to be 98.89% and 100%, respectively, in clinical isolates. The LOD was documented to be 1 parasite/µl at the cut-off ADC value of 20. Parasite density estimated from ADC values showed concordance with microscopically determined parasite density of the cultured P. falciparum 3D7 strain. The LAMP assay performed in our device provides a possible portable platform for its deployment in the point-of-care diagnosis of malaria. Further validation of the quantitative ability of the assay with freshly collected or properly stored clinical samples of known parasitemia is necessary for field applicability. |
| first_indexed | 2024-04-13T18:32:25Z |
| format | Article |
| id | doaj.art-91e4115314104f90b3bbc6da1430fd04 |
| institution | Directory Open Access Journal |
| issn | 2235-2988 |
| language | English |
| last_indexed | 2024-04-13T18:32:25Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Cellular and Infection Microbiology |
| spelling | doaj.art-91e4115314104f90b3bbc6da1430fd042022-12-22T02:35:02ZengFrontiers Media S.A.Frontiers in Cellular and Infection Microbiology2235-29882022-08-011210.3389/fcimb.2022.961832961832Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification deviceMadhu Puri0Harsimran Kaur Brar1Evanka Madan2Rajesh Srinivasan3Kapil Rawat4Sai Siva Gorthi5Geeta Kumari6Raj Sah7Sashi Bhusan Ojha8Subhendu Panigrahi9Gunanidhi Dhangadamajhi10Rohini Muthuswami11Shailja Singh12Rentala Madhubala13School of Life Sciences, Jawaharlal Nehru University, New Delhi, IndiaSchool of Life Sciences, Jawaharlal Nehru University, New Delhi, IndiaSchool of Life Sciences, Jawaharlal Nehru University, New Delhi, IndiaDepartment of Instrumentation and Applied Physics, Indian Institute of Science, Bengaluru, IndiaDepartment of Instrumentation and Applied Physics, Indian Institute of Science, Bengaluru, IndiaDepartment of Instrumentation and Applied Physics, Indian Institute of Science, Bengaluru, IndiaSpecial Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, IndiaSpecial Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, IndiaDepartment of Biotechnology, Maharaja Sriram Chandra Bhanjadeo University, Baripada, IndiaDepartment of General Medicine, VSS Medical College and Hospital, Sambalpur, IndiaDepartment of Biotechnology, Maharaja Sriram Chandra Bhanjadeo University, Baripada, IndiaSchool of Life Sciences, Jawaharlal Nehru University, New Delhi, IndiaSpecial Centre for Molecular Medicine, Jawaharlal Nehru University, New Delhi, IndiaSchool of Life Sciences, Jawaharlal Nehru University, New Delhi, IndiaLAMP diagnosis of malaria is simple and cost-effective with acceptable sensitivity and specificity as compared to standard diagnostic modules such as microscopy, RDTs and nested PCR, and thus its deployment for onsite screening of malaria in resource-limited regions is under consideration. However, the requirement of an electricity-operated dry bath and bulky read-out unit is still a major concern. In an effort to simplify this limitation, we have developed a portable LAMP device and fluorescence readout unit which can be used in the rapid point-of-care diagnosis of malaria. We have developed a point-of-care diagnostic LAMP device that is easy to operate by a mobile application, and the results can be quantified with a fluorescent readout unit. The diagnostic performance of the device was evaluated in 90 P. falciparum-infected clinical isolates stored at 4°C for 6-7 years and 10 freshly collected isolates from healthy volunteers. The LOD and quantitative ability of LAMP in estimating parasitemia levels were revealed with laboratory-grown P. falciparum strain (3D7). The LAMP assay performed in our device was exclusive for P. falciparum detection with sensitivity and specificity determined to be 98.89% and 100%, respectively, in clinical isolates. The LOD was documented to be 1 parasite/µl at the cut-off ADC value of 20. Parasite density estimated from ADC values showed concordance with microscopically determined parasite density of the cultured P. falciparum 3D7 strain. The LAMP assay performed in our device provides a possible portable platform for its deployment in the point-of-care diagnosis of malaria. Further validation of the quantitative ability of the assay with freshly collected or properly stored clinical samples of known parasitemia is necessary for field applicability.https://www.frontiersin.org/articles/10.3389/fcimb.2022.961832/fullLAMPplasmodium falciparumdiagnosisLODparasite densityADC value |
| spellingShingle | Madhu Puri Harsimran Kaur Brar Evanka Madan Rajesh Srinivasan Kapil Rawat Sai Siva Gorthi Geeta Kumari Raj Sah Sashi Bhusan Ojha Subhendu Panigrahi Gunanidhi Dhangadamajhi Rohini Muthuswami Shailja Singh Rentala Madhubala Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device Frontiers in Cellular and Infection Microbiology LAMP plasmodium falciparum diagnosis LOD parasite density ADC value |
| title | Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device |
| title_full | Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device |
| title_fullStr | Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device |
| title_full_unstemmed | Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device |
| title_short | Rapid diagnosis of Plasmodium falciparum malaria using a point-of-care loop-mediated isothermal amplification device |
| title_sort | rapid diagnosis of plasmodium falciparum malaria using a point of care loop mediated isothermal amplification device |
| topic | LAMP plasmodium falciparum diagnosis LOD parasite density ADC value |
| url | https://www.frontiersin.org/articles/10.3389/fcimb.2022.961832/full |
| work_keys_str_mv | AT madhupuri rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT harsimrankaurbrar rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT evankamadan rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT rajeshsrinivasan rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT kapilrawat rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT saisivagorthi rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT geetakumari rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT rajsah rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT sashibhusanojha rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT subhendupanigrahi rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT gunanidhidhangadamajhi rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT rohinimuthuswami rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT shailjasingh rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice AT rentalamadhubala rapiddiagnosisofplasmodiumfalciparummalariausingapointofcareloopmediatedisothermalamplificationdevice |