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1دورية أكاديمية
المؤلفون: Hossain, Faria, Picado, Albert, Owen, Sophie I., Ghosh, Prakash, Chowdhury, Rajashree, Maruf, Shomik, Ashfaq Khan, Md. Anik, Rashid, Md. Utba, Nath, Rupen, Baker, James, Ghosh, Debashis, Adams, Emily R., Duthie, Malcolm S., Hossain, Md. Sakhawat, Basher, Ariful, Nath, Proggananda, Aktar, Fatima, Cruz, Israel, Mondal, Dinesh
مصطلحات موضوعية: visceral leishmaniasis, diagnosis, LAMP, qPCR, urine ELISA, elimination, diagnostics for VL postelimination era, info:eu-repo/classification/ddc/610, ddc:610
الوصف: With reduced prevalence of visceral leishmaniasis (VL) in the Indian subcontinent (ISC), direct and field deployable diagnostic tests are needed to implement an effective diagnostic and surveillance algorithm for post-elimination VL control. In this regard, here we investigated the diagnostic efficacies of a loop-mediated isothermal amplification (LAMP) assay (Loopamp™ Leishmania Detection Kit, Eiken Chemical CO., Ltd, Japan), a real-time quantitative PCR assay (qPCR) and the Leishmania antigen ELISA (CLIN-TECH, UK) with different sampling techniques and evaluated their prospect to incorporate into post-elimination VL control strategies. Eighty clinically and rK39 rapid diagnostic test confirmed VL cases and 80 endemic healthy controls were enrolled in the study. Peripheral blood and dried blood spots (DBS) were collected from all the participants at the time of diagnosis. DNA was extracted from whole blood (WB) and DBS via silica columns (QIAGEN) and boil & spin (B&S) methods and tested with qPCR and Loopamp. Urine was collected from all participants at the time of diagnosis and was directly subjected to the Leishmania antigen ELISA. 41 patients were followed up and urine samples were collected at day 30 and day 180 after treatment and ELISA was performed. The sensitivities of the Loopamp-WB(B&S) and Loopamp-WB(QIA) were 96.2% (95% CI 89·43-99·22) and 95% (95% CI 87·69-98·62) respectively. The sensitivity of Loopamp- DBS(QIA) was 85% (95% CI 75·26- 92·00). The sensitivities of the qPCR-WB(QIA) and qPCR-DBS(QIA) were 93.8% (95% CI 86·01-97·94) and 72.5% (95% CI 61·38-81·90) respectively. The specificity of all molecular assays was 100%. The sensitivity and specificity of the Leishmania antigen ELISA were 97.5% (95% CI 91·47-99·70) and 91.95% (95% CI 84·12-96·70) respectively. The Leishmania antigen ELISA depicted clinical cure at day 180 in all the followed-up cases. Efficacy and sustainability identify the Loopamp-WB(B&S) and the Leishmania antigen ELISA as promising and minimally invasive VL diagnostic tools to support VL diagnostic and surveillance activities respectively in the post-elimination era.
العلاقة: 670759
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2دورية أكاديمية
المؤلفون: Rock, Kat S, Chapman, Lloyd A C, Dobson, Andrew P, Adams, Emily R, Hollingsworth, T Déirdre
الوصف: Background Neglected tropical diseases are responsible for considerable morbidity and mortality in low-income populations. International efforts have reduced their global burden, but transmission is persistent and case-finding-based interventions rarely target asymptomatic individuals. Methods We develop a generic mathematical modeling framework for analyzing the dynamics of visceral leishmaniasis in the Indian sub-continent (VL), gambiense sleeping sickness (gHAT), and Chagas disease and use it to assess the possible contribution of asymptomatics who later develop disease (pre-symptomatics) and those who do not (non-symptomatics) to the maintenance of infection. Plausible interventions, including active screening, vector control, and reduced time to detection, are simulated for the three diseases.ResultsWe found that the high asymptomatic contribution to transmission for Chagas and gHAT and the apparently high basic reproductive number of VL may undermine long-term control. However, the ability to treat some asymptomatics for Chagas and gHAT should make them more controllable, albeit over relatively long time periods due to the slow dynamics of these diseases. For VL, the toxicity of available therapeutics means the asymptomatic population cannot currently be treated, but combining treatment of symptomatics and vector control could yield a quick reduction in transmission.ConclusionsDespite the uncertainty in natural history, it appears there is already a relatively good toolbox of interventions to eliminate gHAT, and it is likely that Chagas will need improvements to diagnostics and their use to better target pre-symptomatics. The situation for VL is less clear, and model predictions could be improved by additional empirical data. However, interventions may have to improve to successfully eliminate this disease.
وصف الملف: text
العلاقة: https://eprints.lancs.ac.uk/id/eprint/219379/1Test/; https://eprints.lancs.ac.uk/id/eprint/219379/3/ciae096.pdfTest; Rock, Kat S and Chapman, Lloyd A C and Dobson, Andrew P and Adams, Emily R and Hollingsworth, T Déirdre (2024) The Hidden Hand of Asymptomatic Infection Hinders Control of Neglected Tropical Diseases : A Modeling Analysis. Clinical infectious diseases : an official publication of the Infectious Diseases Society of America, 78 (Suppl.). S175-S182. ISSN 1058-4838
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3دورية أكاديمية
المؤلفون: Boonyuen, Usa, Jacob, Beatriz Aira C., Wongwigkan, Jutamas, Chamchoy, Kamonwan, Singha-art, Natsamon, Pengsuk, Natnicha, Songdej, Duantida, Adams, Emily R., Edwards, Thomas, Chamnanchanunt, Supat, Amran, Syazwani Itri, Latif, Nurriza Ab, Louis, Naveen Eugene, Chandran, Shamini
المساهمون: Specific League Funds 2023 from Mahidol University, Ministry of Health – Kingdom of Saudi Arabia, Mahidol University
المصدر: Malaria Journal ; volume 23, issue 1 ; ISSN 1475-2875
مصطلحات موضوعية: Infectious Diseases, Parasitology
الوصف: Background It was hypothesized that glucose-6-phosphate dehydrogenase (G6PD) deficiency confers a protective effect against malaria infection, however, safety concerns have been raised regarding haemolytic toxicity caused by radical cure with 8-aminoquinolines in G6PD-deficient individuals. Malaria elimination and control are also complicated by the high prevalence of G6PD deficiency in malaria-endemic areas. Hence, accurate identification of G6PD deficiency is required to identify those who are eligible for malaria treatment using 8-aminoquinolines. Methods The prevalence of G6PD deficiency among 408 Thai participants diagnosed with malaria by microscopy (71), and malaria-negative controls (337), was assessed using a phenotypic test based on water-soluble tetrazolium salts. High-resolution melting (HRM) curve analysis was developed from a previous study to enable the detection of 15 common missense, synonymous and intronic G6PD mutations in Asian populations. The identified mutations were subjected to biochemical and structural characterisation to understand the molecular mechanisms underlying enzyme deficiency. Results Based on phenotypic testing, the prevalence of G6PD deficiency (< 30% activity) was 6.13% (25/408) and intermediate deficiency (30–70% activity) was found in 15.20% (62/408) of participants. Several G6PD genotypes with newly discovered double missense variants were identified by HRM assays, including G6PD Gaohe + Viangchan, G6PD Valladolid + Viangchan and G6PD Canton + Viangchan. A significantly high frequency of synonymous (c.1311C>T) and intronic (c.1365-13T>C and c.486-34delT) mutations was detected with intermediate to normal enzyme activity. The double missense mutations were less catalytically active than their corresponding single missense mutations, resulting in severe enzyme deficiency. While the mutations had a minor effect on binding affinity, structural instability was a key contributor to the enzyme deficiency observed in G6PD-deficient individuals. Conclusions ...
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4دورية أكاديمية
المؤلفون: Toor, Jaspreet, Adams, Emily R, Aliee, Maryam, Amoah, Benjamin, Anderson, Roy M, Ayabina, Diepreye, Bailey, Robin, Basáñez, Maria-Gloria, Blok, David J, Blumberg, Seth, Borlase, Anna, Rivera, Rocio Caja, Castaño, María Soledad, Chitnis, Nakul, Coffeng, Luc E, Crump, Ronald E, Das, Aatreyee, Davis, Christopher N, Davis, Emma L, Deiner, Michael S, Diggle, Peter J, Fronterre, Claudio, Giardina, Federica, Giorgi, Emanuele, Graham, Matthew, Hamley, Jonathan ID, Huang, Ching-I, Kura, Klodeta, Lietman, Thomas M, Lucas, Tim CD, Malizia, Veronica, Medley, Graham F, Meeyai, Aronrag, Michael, Edwin, Porco, Travis C, Prada, Joaquin M, Rock, Kat S, Le Rutte, Epke A, Smith, Morgan E, Spencer, Simon EF, Stolk, Wilma A, Touloupou, Panayiota, Vasconcelos, Andreia, Vegvari, Carolin, de Vlas, Sake J, Walker, Martin, Hollingsworth, T Déirdre
المصدر: Clinical Infectious Diseases. 72(8)
مصطلحات موضوعية: Rare Diseases, Infectious Diseases, Vector-Borne Diseases, Orphan Drug, Infection, Good Health and Well Being, COVID-19, Humans, Neglected Diseases, Pandemics, SARS-CoV-2, Tropical Medicine, neglected tropical diseases, coronavirus, modeling, Biological Sciences, Medical and Health Sciences, Microbiology
الوصف: Due to the COVID-19 pandemic, many key neglected tropical disease (NTD) activities have been postponed. This hindrance comes at a time when the NTDs are progressing towards their ambitious goals for 2030. Mathematical modelling on several NTDs, namely gambiense sleeping sickness, lymphatic filariasis, onchocerciasis, schistosomiasis, soil-transmitted helminthiases (STH), trachoma, and visceral leishmaniasis, shows that the impact of this disruption will vary across the diseases. Programs face a risk of resurgence, which will be fastest in high-transmission areas. Furthermore, of the mass drug administration diseases, schistosomiasis, STH, and trachoma are likely to encounter faster resurgence. The case-finding diseases (gambiense sleeping sickness and visceral leishmaniasis) are likely to have fewer cases being detected but may face an increasing underlying rate of new infections. However, once programs are able to resume, there are ways to mitigate the impact and accelerate progress towards the 2030 goals.
وصف الملف: application/pdf
الوصول الحر: https://escholarship.org/uc/item/6986r862Test
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5دورية أكاديمية
المؤلفون: Palomino-Padilla, Sandra, Finch, Lorna, de Vos, Margaretha, Savage, Helen, Villa-Castillo, Luz, Hayward, Gail, Cook, Eloïse, Escadafal, Camille, Body, Richard, Adams, Emily R., Ugarte-Gil, Cesar, CubasAtienzar, Ana
المصدر: PLoS ONE, Vol 18, Issue 3, e0281925.
مصطلحات موضوعية: QW 573 Antigens, WC 506 COVID-19
الوصف: Objectives: In order to generate independent performance data regarding accuracy of COVID-19 antigen-based rapid diagnostic tests (Ag-RDTs), prospective diagnostic evaluation studies across multiple sites are required to evaluate their performance in different clinical settings. This report describes the clinical evaluation the GENEDIA W COVID-19 Ag Device (Green Cross Medical Science Corp., Chungbuk, Korea) and the ActiveXpress+ COVID-19 Complete Testing Kit (Edinburgh Genetics Ltd, UK), in two testing sites Peru and the United Kingdom. Methods: Nasopharyngeal swabs collected from 456 symptomatic patients at primary points of care in Lima, Peru and 610 symptomatic participants at a COVID-19 Drive-Through testing site in Liverpool, England were analyzed by Ag-RDT and compared to RT-PCR. Analytical evaluation of both Ag-RDTs was assessed using serial dilutions of direct culture supernatant of a clinical SARS-CoV-2 isolate from the B.1.1.7 lineage. Results: For GENEDIA brand, the values of overall sensitivity and specificity were 60.4% [95% CI 52.4–67.9%], and 99.2% [95% CI 97.6–99.7%] respectively; and for Active Xpress+ the overall values of sensitivity and specificity were 66.2% [95% CI 54.0–76.5%], and 99.6% [95% CI 97.9–99.9%] respectively. The analytical limit of detection was determined at 5.0 x 102 pfu/ml what equals to approximately 1.0 x 104 gcn/ml for both Ag-RDTs. The UK cohort had lower median Ct values compared to that of Peru during both evaluations. When split by Ct, both Ag-RDTs had optimum sensitivities at Ct<20 (in Peru; 95% [95% CI 76.4–99.1%] and 100.0% [95% CI 74.1–100.0%] and in the UK; 59.2% [95% CI 44.2–73.0%] and 100.0% [95% CI 15.8–100.0%], for the GENDIA and the ActiveXpress+, respectively). Conclusions: Whilst the overall clinical sensitivity of the Genedia did not meet WHO minimum performance requirements for rapid immunoassays in either cohort, the ActiveXpress+ did so for the small UK cohort. This study illustrates comparative performance of Ag-RDTs across two global settings and ...
وصف الملف: text
العلاقة: https://archive.lstmed.ac.uk/22072/1/pone.0281925.pdfTest; Palomino-Padilla, Sandra, Finch, Lorna, de Vos, Margaretha, Savage, Helen, Villa-Castillo, Luz, Hayward, Gail, Cook, Eloïse, Escadafal, Camille, Body, Richard, Adams, Emily R., Ugarte-Gil, Cesar and CubasAtienzar, Ana (2023) 'Diagnostic performance of GENEDIA W and ActiveXpress+ COVID-19 antigens tests among symptomatic individuals in Peru and The United Kingdom'. PLoS ONE, Vol 18, Issue 3, e0281925.
الإتاحة: https://doi.org/10.1371/journal.pone.0281925Test
https://archive.lstmed.ac.uk/22072Test/
https://archive.lstmed.ac.uk/22072/1/pone.0281925.pdfTest -
6دورية أكاديمية
المؤلفون: Greenland-Bews, Caitlin, Byrne, Rachel L., Owen, Sophie I., Watkins, Rachel L., Bengey, Daisy, Buist, Kate, Clerkin, Karina, Escadafal, Camille, Finch, Lorna S., Gould, Susan, Giorgi, Emanuele, Hodgkinson, Andy, Mashenko, Larysa, Powell, Darren, Savage, Helen R., Thompson, Caitlin R., Turtle, Lance, Wardale, Jahanara, Wooding, Dominic, Edwards, Thomas, Atienzar, Ana Cubas, Adams, Emily R.
المصدر: BMC Infectious Diseases, Vol 23, Issue 1, e110.
مصطلحات موضوعية: QS 4 General works. Classify here works on regional anatomy, QW 575 Antibodies, QW 806 Vaccination
الوصف: Background: Rapid determination of an individual's antibody status can be beneficial in understanding an individual's immune response to SARS-CoV-2 and for initiation of therapies that are only deemed effective in sero-negative individuals. Antibody lateral flow tests (LFTs) have potential to address this need as a rapid, point of care test. Methods: Here we present a proof-of-concept evaluation of eight LFT brands using sera from 95 vaccinated individuals to determine sensitivity for detecting vaccination generated antibodies. Samples were analysed on eight different brands of antibody LFT and an automated chemiluminescent microparticle immunoassay (CMIA) that identifies anti-spike antibodies which was used as our reference standard. Results: All 95 (100%) participants tested positive for anti-spike antibodies by the chemiluminescent microparticle immunoassay (CMIA) reference standard post-dose two of their SARS-CoV-2 vaccine: BNT162b2 (Pfizer/BioNTech, n = 60), AZD1222 (AstraZeneca, n = 31), mRNA-1273 (Moderna, n = 2) and Undeclared Vaccine Brand (n = 2). Sensitivity increased from dose one to dose two in six out of eight LFTs with three tests achieving 100% sensitivity at dose two in detecting anti-spike antibodies. Conclusions: These tests are demonstrated to be highly sensitive to detect raised antibody levels in vaccinated individuals. RDTs are low cost and rapid alternatives to ELISA based systems.
وصف الملف: text
العلاقة: https://archive.lstmed.ac.uk/22054/1/s12879-023-08033-1.pdfTest; Greenland-Bews, Caitlin, Byrne, Rachel L., Owen, Sophie I. orcid:0000-0002-0458-2357 , Watkins, Rachel L., Bengey, Daisy, Buist, Kate, Clerkin, Karina, Escadafal, Camille, Finch, Lorna S., Gould, Susan, Giorgi, Emanuele, Hodgkinson, Andy, Mashenko, Larysa, Powell, Darren, Savage, Helen R., Thompson, Caitlin R., Turtle, Lance, Wardale, Jahanara, Wooding, Dominic, Edwards, Thomas, Atienzar, Ana Cubas and Adams, Emily R. orcid:0000-0002-0816-2835 (2023) 'Evaluation of eight lateral flow tests for the detection of anti-SARS-CoV-2 antibodies in a vaccinated population'. BMC Infectious Diseases, Vol 23, Issue 1, e110.
الإتاحة: https://doi.org/10.1186/s12879-023-08033-1Test
https://archive.lstmed.ac.uk/22054Test/
https://archive.lstmed.ac.uk/22054/1/s12879-023-08033-1.pdfTest -
7دورية أكاديمية
المؤلفون: Edwards, Thomas, Williams, Christopher T., Olwala, Macrine, Andang’o, Pauline, Otieno, Walter, Nalwa, Grace N., Akindolire, Abimbola, Cubas-Atienzar, Ana I., Ross, Toby, Tongo, Olukemi O., Adams, Emily R., Nabwera, Helen, Allen, Stephen
المصدر: Antimicrobial Resistance & Infection Control, Vol 12, Issue 1.
مصطلحات موضوعية: QS 4 General works. Classify here works on regional anatomy, WY 157.3 Maternal-child nursing. Neonatal nursing. Perinatal nursing
الوصف: Objectives: Neonatal sepsis, a major cause of death amongst infants in sub-Saharan Africa, is often gut derived. Gut colonisation by Enterobacteriaceae producing extended spectrum beta-lactamase (ESBL) or carbapenemase enzymes can lead to antimicrobial-resistant (AMR) or untreatable infections. We sought to explore the rates of colonisation by ESBL or carbapenemase producers in two neonatal units (NNUs) in West and East Africa. Methods: Stool and rectal swab samples were taken at multiple timepoints from newborns admitted to the NNUs at the University College Hospital, Ibadan, Nigeria and the Jaramogi Oginga Odinga Teaching and Referral Hospital, Kisumu, western Kenya. Samples were tested for ESBL and carbapenemase genes using a previously validated qPCR assay. Kaplan-Meier survival analysis was used to examine colonisation rates at both sites. Results: In total 119 stool and rectal swab samples were taken from 42 infants admitted to the two NNUs. Colonisation with ESBL (37 infants, 89%) was more common than with carbapenemase producers (26, 62.4%; P = 0.093). Median survival time before colonisation with ESBL organisms was 7 days and with carbapenemase producers 16 days (P = 0.035). The majority of ESBL genes detected belonged to the CTX-M-1 (36/38; 95%), and CTX-M-9 (2/36; 5%) groups, and the most prevalent carbapenemase was blaNDM (27/29, 93%). Conclusions: Gut colonisation of neonates by AMR organisms was common and occurred rapidly in NNUs in Kenya and Nigeria. Active surveillance of colonisation will improve the understanding of AMR in these settings and guide infection control and antibiotic prescribing practice to improve clinical outcomes.
وصف الملف: text
العلاقة: https://archive.lstmed.ac.uk/22053/9/s13756-023-01216-0.pdfTest; Edwards, Thomas, Williams, Christopher T., Olwala, Macrine, Andang’o, Pauline, Otieno, Walter, Nalwa, Grace N., Akindolire, Abimbola, Cubas-Atienzar, Ana I., Ross, Toby, Tongo, Olukemi O., Adams, Emily R. orcid:0000-0002-0816-2835 , Nabwera, Helen and Allen, Stephen (2023) 'Molecular surveillance reveals widespread colonisation by carbapenemase and extended spectrum beta-lactamase producing organisms in neonatal units in Kenya and Nigeria'. Antimicrobial Resistance & Infection Control, Vol 12, Issue 1.
الإتاحة: https://doi.org/10.1186/s13756-023-01216-0Test
https://archive.lstmed.ac.uk/22053Test/
https://archive.lstmed.ac.uk/22053/9/s13756-023-01216-0.pdfTest -
8دورية أكاديمية
المؤلفون: Fraser, Alice J., Greenland-Bews, Caitlin, Kelly, Daniel, Williams, Christopher T., Body, Rick, Adams, Emily R., Cubas Atienzar, Ana, Edwards, Thomas, Allen, David J., Dark, Paul
المصدر: Fraser , A J , Greenland-Bews , C , Kelly , D , Williams , C T , CONDOR Steering Group , LSTM Diagnostics Group , Body , R , Adams , E R , Cubas Atienzar , A , Edwards , T , Allen , D J & Dark , P 2023 , ' A high-resolution melt curve toolkit to identify lineage-defining SARS-CoV-2 mutations ' , Scientific Reports , vol. 13 , no. 1 , 3887 . https://doi.org/10.1038/s41598-023-30754-1Test
مصطلحات موضوعية: Humans, COVID-19, SARS-CoV-2/genetics, Mutation, Biological Assay, Genomics
الوصف: The emergence of severe acute respiratory syndrome 2 (SARS-CoV-2) variants of concern (VOCs), with mutations linked to increased transmissibility, vaccine escape and virulence, has necessitated the widespread genomic surveillance of SARS-CoV-2. This has placed a strain on global sequencing capacity, especially in areas lacking the resources for large scale sequencing activities. Here we have developed three separate multiplex high-resolution melting assays to enable the identification of Alpha, Beta, Delta and Omicron VOCs. The assays were evaluated against whole genome sequencing on upper-respiratory swab samples collected during the Alpha, Delta and Omicron [BA.1] waves of the UK pandemic. The sensitivities of the eight individual primer sets were all 100%, and specificity ranged from 94.6 to 100%. The multiplex HRM assays have potential as a tool for high throughput surveillance of SARS-CoV-2 VOCs, particularly in areas with limited genomics facilities.
الإتاحة: https://doi.org/10.1038/s41598-023-30754-1Test
https://research.manchester.ac.uk/en/publications/cb2a3e5b-e457-4982-825a-92489ca3016eTest
http://www.scopus.com/inward/record.url?scp=85150228906&partnerID=8YFLogxKTest -
9دورية أكاديمية
المؤلفون: Ding, Xavier C., Incardona, Sandra, Serra-Casas, Elisa, Charnaud, Sarah C., Slater, Hannah C., Domingo, Gonzalo J., Adams, Emily R., ter Kuile, Feiko O., Samuels, Aaron M., Kariuki, Simon, Dittrich, Sabine
المصدر: Malaria Journal ; volume 22, issue 1 ; ISSN 1475-2875
مصطلحات موضوعية: Infectious Diseases, Parasitology
الوصف: Background Rapid diagnostic tests (RDTs) are effective tools to diagnose and inform the treatment of malaria in adults and children. The recent development of a highly sensitive rapid diagnostic test (HS-RDT) for Plasmodium falciparum has prompted questions over whether it could improve the diagnosis of malaria in pregnancy and pregnancy outcomes in malaria endemic areas. Methods This landscape review collates studies addressing the clinical performance of the HS-RDT. Thirteen studies were identified comparing the HS-RDT and conventional RDT (co-RDT) to molecular methods to detect malaria in pregnancy. Using data from five completed studies, the association of epidemiological and pregnancy-related factors on the sensitivity of HS-RDT, and comparisons with co-RDT were investigated. The studies were conducted in 4 countries over a range of transmission intensities in largely asymptomatic women. Results Sensitivity of both RDTs varied widely (HS-RDT range 19.6 to 85.7%, co-RDT range 22.8 to 82.8% compared to molecular testing) yet HS-RDT detected individuals with similar parasite densities across all the studies including different geographies and transmission areas [geometric mean parasitaemia around 100 parasites per µL (p/µL)]. HS-RDTs were capable of detecting low-density parasitaemias and in one study detected around 30% of infections with parasite densities of 0–2 p/µL compared to the co-RDT in the same study which detected around 15%. Conclusion The HS-RDT has a slightly higher analytical sensitivity to detect malaria infections in pregnancy than co-RDT but this mostly translates to only fractional and not statistically significant improvement in clinical performance by gravidity, trimester, geography or transmission intensity. The analysis presented here highlights the need for larger and more studies to evaluate incremental improvements in RDTs. The HS-RDT could be used in any situation where co-RDT are currently used for P. falciparum diagnosis, if storage conditions can be adhered to.
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10دورية أكاديمية
المؤلفون: Chamchoy, Kamonwan, Sudsumrit, Sirapapha, Wongwigkan, Jutamas, Petmitr, Songsak, Songdej, Duantida, Adams, Emily R., Edwards, Thomas, Leartsakulpanich, Ubolsree, Boonyuen, Usa
المساهمون: Bancone, Germana, Chulabhorn Royal Academy, Mahidol University (Fundamental Fund: fiscal year 2023 by National Science Research and Innovation Fund
المصدر: PLOS ONE ; volume 18, issue 11, page e0294200 ; ISSN 1932-6203
الوصف: Glucose-6-phosphate dehydrogenase (G6PD) deficiency is an X-linked enzymopathy caused by mutations in the G6PD gene. A medical concern associated with G6PD deficiency is acute hemolytic anemia induced by certain foods, drugs, and infections. Although phenotypic tests can correctly identify hemizygous males, as well as homozygous and compound heterozygous females, heterozygous females with a wide range of G6PD activity may be misclassified as normal. This study aimed to develop multiplex high-resolution melting (HRM) analyses to enable the accurate detection of G6PD mutations, especially among females with heterozygous deficiency. Multiplex HRM assays were developed to detect six G6PD variants, i.e., G6PD Gaohe (c.95A>G), G6PD Chinese-4 (c.392G>T), G6PD Mahidol (c.487G>A), G6PD Viangchan (c.871G>A), G6PD Chinese-5 (c.1024C>T), and G6PD Union (c.1360C>T) in two reactions. The assays were validated and then applied to genotype G6PD mutations in 248 Thai females. The sensitivity of the HRM assays developed was 100% [95% confidence interval (CI): 94.40%–100%] with a specificity of 100% (95% CI: 88.78%–100%) for detecting these six mutations. The prevalence of G6PD deficiency was estimated as 3.63% (9/248) for G6PD deficiency and 31.05% (77/248) for intermediate deficiency by phenotypic assay. The developed HRM assays identified three participants with normal enzyme activity as heterozygous for G6PD Viangchan. Interestingly, a deletion in intron 5 nucleotide position 637/638 (c.486-34delT) was also detected by the developed HRM assays. G6PD genotyping revealed a total of 12 G6PD genotypes, with a high prevalence of intronic variants. Our results suggested that HRM analysis-based genotyping is a simple and reliable approach for detecting G6PD mutations, and could be used to prevent the misdiagnosis of heterozygous females by phenotypic assay. This study also sheds light on the possibility of overlooking intronic variants, which could affect G6PD expression and contribute to enzyme deficiency.
النص الكامل