{"title":"评估和验证基于 PrintrLab 的 LAMP 检测方法,以确定玻利维亚新生儿中的克鲁斯锥虫:概念验证研究。","authors":"","doi":"10.1016/S2666-5247(24)00110-1","DOIUrl":null,"url":null,"abstract":"<div><h3>Background</h3><p>Vertical transmission of <em>Trypanosoma cruzi</em> represents approximately 20% of new Chagas disease cases. Early detection and treatment for women of childbearing age and newborns is a public health priority, but the lack of a simple and reliable diagnostic test remains a major barrier. We aimed to evaluate the performance of a point-of-care loop-mediated isothermal amplification (LAMP) assay for the detection of <em>T cruzi</em>.</p></div><div><h3>Methods</h3><p>In this proof-of-concept study, we coupled a low-cost 3D printer repurposed for sample preparation and amplification (PrintrLab) to the Eiken <em>T cruzi-</em>LAMP prototype to detect vertically transmitted <em>T cruzi</em>, which we compared with standardised PCR and with the gold-standard algorithm (microscopy at birth and 2 months and serological study several months later). We screened pregnant women from two hospitals in the Bolivian Gran Chaco province, and those who were seropositive for <em>T cruzi</em> were offered the opportunity for their newborns to be enrolled in the study. Newborns were tested by microscopy, LAMP, and PCR at birth and 2 months, and by serology at 8 months.</p></div><div><h3>Findings</h3><p>Between April 23 and Nov 17, 2018, 986 mothers were screened, among whom 276 were seropositive for <em>T cruzi</em> (28·0% prevalence, 95% CI 25·6–31·2). In total, 224 infants born to 221 seropositive mothers completed 8 months of follow-up. Congenital transmission was detected in nine of the 224 newborns (4·0% prevalence, 1·9–7·5) by direct microscopy observation, and 14 more cases were diagnosed serologically (6·3%, 3·6–10·3), accounting for an overall vertical transmission rate of 10·3% (6·6–15·0; 23 of 224). All microscopy-positive newborns were positive by PrintrLab-LAMP and by PCR, while these techniques respectively detected four and five extra positive cases among the remaining 215 microscopy-negative newborns.</p></div><div><h3>Interpretation</h3><p>The PrintrLab-LAMP yielded a higher sensitivity than microscopy-based analysis. Considering the simpler use and expected lower cost of LAMP compared with PCR, our findings encourage its evaluation in a larger study over a wider geographical area.</p></div><div><h3>Funding</h3><p>Inter-American Development Bank.</p></div>","PeriodicalId":46633,"journal":{"name":"Lancet Microbe","volume":null,"pages":null},"PeriodicalIF":20.9000,"publicationDate":"2024-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S2666524724001101/pdfft?md5=e5fdd2cd8e083b58c863871674c0eec7&pid=1-s2.0-S2666524724001101-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Evaluation and validation of a PrintrLab-based LAMP assay to identify Trypanosoma cruzi in newborns in Bolivia: a proof-of-concept study\",\"authors\":\"\",\"doi\":\"10.1016/S2666-5247(24)00110-1\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Background</h3><p>Vertical transmission of <em>Trypanosoma cruzi</em> represents approximately 20% of new Chagas disease cases. Early detection and treatment for women of childbearing age and newborns is a public health priority, but the lack of a simple and reliable diagnostic test remains a major barrier. We aimed to evaluate the performance of a point-of-care loop-mediated isothermal amplification (LAMP) assay for the detection of <em>T cruzi</em>.</p></div><div><h3>Methods</h3><p>In this proof-of-concept study, we coupled a low-cost 3D printer repurposed for sample preparation and amplification (PrintrLab) to the Eiken <em>T cruzi-</em>LAMP prototype to detect vertically transmitted <em>T cruzi</em>, which we compared with standardised PCR and with the gold-standard algorithm (microscopy at birth and 2 months and serological study several months later). We screened pregnant women from two hospitals in the Bolivian Gran Chaco province, and those who were seropositive for <em>T cruzi</em> were offered the opportunity for their newborns to be enrolled in the study. Newborns were tested by microscopy, LAMP, and PCR at birth and 2 months, and by serology at 8 months.</p></div><div><h3>Findings</h3><p>Between April 23 and Nov 17, 2018, 986 mothers were screened, among whom 276 were seropositive for <em>T cruzi</em> (28·0% prevalence, 95% CI 25·6–31·2). In total, 224 infants born to 221 seropositive mothers completed 8 months of follow-up. Congenital transmission was detected in nine of the 224 newborns (4·0% prevalence, 1·9–7·5) by direct microscopy observation, and 14 more cases were diagnosed serologically (6·3%, 3·6–10·3), accounting for an overall vertical transmission rate of 10·3% (6·6–15·0; 23 of 224). All microscopy-positive newborns were positive by PrintrLab-LAMP and by PCR, while these techniques respectively detected four and five extra positive cases among the remaining 215 microscopy-negative newborns.</p></div><div><h3>Interpretation</h3><p>The PrintrLab-LAMP yielded a higher sensitivity than microscopy-based analysis. Considering the simpler use and expected lower cost of LAMP compared with PCR, our findings encourage its evaluation in a larger study over a wider geographical area.</p></div><div><h3>Funding</h3><p>Inter-American Development Bank.</p></div>\",\"PeriodicalId\":46633,\"journal\":{\"name\":\"Lancet Microbe\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":20.9000,\"publicationDate\":\"2024-09-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S2666524724001101/pdfft?md5=e5fdd2cd8e083b58c863871674c0eec7&pid=1-s2.0-S2666524724001101-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Lancet Microbe\",\"FirstCategoryId\":\"99\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2666524724001101\",\"RegionNum\":1,\"RegionCategory\":\"生物学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"INFECTIOUS DISEASES\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Lancet Microbe","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2666524724001101","RegionNum":1,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"INFECTIOUS DISEASES","Score":null,"Total":0}
Evaluation and validation of a PrintrLab-based LAMP assay to identify Trypanosoma cruzi in newborns in Bolivia: a proof-of-concept study
Background
Vertical transmission of Trypanosoma cruzi represents approximately 20% of new Chagas disease cases. Early detection and treatment for women of childbearing age and newborns is a public health priority, but the lack of a simple and reliable diagnostic test remains a major barrier. We aimed to evaluate the performance of a point-of-care loop-mediated isothermal amplification (LAMP) assay for the detection of T cruzi.
Methods
In this proof-of-concept study, we coupled a low-cost 3D printer repurposed for sample preparation and amplification (PrintrLab) to the Eiken T cruzi-LAMP prototype to detect vertically transmitted T cruzi, which we compared with standardised PCR and with the gold-standard algorithm (microscopy at birth and 2 months and serological study several months later). We screened pregnant women from two hospitals in the Bolivian Gran Chaco province, and those who were seropositive for T cruzi were offered the opportunity for their newborns to be enrolled in the study. Newborns were tested by microscopy, LAMP, and PCR at birth and 2 months, and by serology at 8 months.
Findings
Between April 23 and Nov 17, 2018, 986 mothers were screened, among whom 276 were seropositive for T cruzi (28·0% prevalence, 95% CI 25·6–31·2). In total, 224 infants born to 221 seropositive mothers completed 8 months of follow-up. Congenital transmission was detected in nine of the 224 newborns (4·0% prevalence, 1·9–7·5) by direct microscopy observation, and 14 more cases were diagnosed serologically (6·3%, 3·6–10·3), accounting for an overall vertical transmission rate of 10·3% (6·6–15·0; 23 of 224). All microscopy-positive newborns were positive by PrintrLab-LAMP and by PCR, while these techniques respectively detected four and five extra positive cases among the remaining 215 microscopy-negative newborns.
Interpretation
The PrintrLab-LAMP yielded a higher sensitivity than microscopy-based analysis. Considering the simpler use and expected lower cost of LAMP compared with PCR, our findings encourage its evaluation in a larger study over a wider geographical area.
期刊介绍:
The Lancet Microbe is a gold open access journal committed to publishing content relevant to clinical microbiologists worldwide, with a focus on studies that advance clinical understanding, challenge the status quo, and advocate change in health policy.