Pub Date : 2025-10-01Epub Date: 2025-08-05DOI: 10.1016/j.jcv.2025.105852
Long Chen, Hong Yang, Hai-Long Zhang, Xiao-Lu Shi, Bo Peng, Jun Meng
Objectives: To investigate the etiological characteristics of acute conjunctivitis in Shenzhen, China.
Methods: A total of 1234 conjunctival swabs collected between 2018 and 2024 were examined for coxsackievirus A24 variant (CVA24v), enterovirus D70 (EV-D70) and human adenovirus (HAdV). Complete VP1 sequences of CVA24v strains were determined and analyzed. HAdV was genotyped by PCR methods targeting the three genes (penton base, hexon and fiber) and sequencing. SPSS 22.0 software was used for statistical analysis.
Results: CVA24v was first detected in 2023, with a detection rate of 33.3 % (63/189). No EV-D70 was detected in 2018-2024. The annual distributions of HAdV-infected patients were 52.6 % (101/192), 62.7 % (111/177), 21.1 % (37/175), 9.0 % (14/155), 10.5 % (17/162), 15.9 % (30/189) and 21.7 % (40/184), respectively. CVA24v strains from this study clustered in a clade with significant temporal aggregation characteristic within the genotype GIV. Eight amino acid variation sites (T11A, I16L, K20I, L32P, K105R, A146T, R277G and P280S) were observed in VP1 sequences of CVA24v strains from this study when compared to the close strains. Conjunctivitis patients with one of the three symptoms (weakness, fever and blurred vision) had a higher detection rate of HAdV. Eleven known HAdV genotypes (HAdV-B3, -B14, -B21, HAdV-D8, -D37, -D42, -D53, -D64, -D85, -D115 and HAdV-E4) and 7 unknown genotypes were detected in 2020-2024, with HAdV-D37 (30.9 %), HAdV-D115 (21.1 %) and HAdV-B3 (17.1 %) being the three predominant genotypes.
Conclusions: The unique genetic characteristics were observed in Shenzhen CVA24v strains. HAdVs associated with conjunctivitis exhibited a high degree of genotypic diversity in Shenzhen, and HAdV-D115 related to conjunctivitis was first reported in this study.
{"title":"Pathogen surveillance of acute conjunctivitis reveals recent emergence of coxsackievirus A24 variants and high genotypic diversity of human adenoviruses in Shenzhen, China, 2018-2024.","authors":"Long Chen, Hong Yang, Hai-Long Zhang, Xiao-Lu Shi, Bo Peng, Jun Meng","doi":"10.1016/j.jcv.2025.105852","DOIUrl":"10.1016/j.jcv.2025.105852","url":null,"abstract":"<p><strong>Objectives: </strong>To investigate the etiological characteristics of acute conjunctivitis in Shenzhen, China.</p><p><strong>Methods: </strong>A total of 1234 conjunctival swabs collected between 2018 and 2024 were examined for coxsackievirus A24 variant (CVA24v), enterovirus D70 (EV-D70) and human adenovirus (HAdV). Complete VP1 sequences of CVA24v strains were determined and analyzed. HAdV was genotyped by PCR methods targeting the three genes (penton base, hexon and fiber) and sequencing. SPSS 22.0 software was used for statistical analysis.</p><p><strong>Results: </strong>CVA24v was first detected in 2023, with a detection rate of 33.3 % (63/189). No EV-D70 was detected in 2018-2024. The annual distributions of HAdV-infected patients were 52.6 % (101/192), 62.7 % (111/177), 21.1 % (37/175), 9.0 % (14/155), 10.5 % (17/162), 15.9 % (30/189) and 21.7 % (40/184), respectively. CVA24v strains from this study clustered in a clade with significant temporal aggregation characteristic within the genotype GIV. Eight amino acid variation sites (T11A, I16L, K20I, L32P, K105R, A146T, R277G and P280S) were observed in VP1 sequences of CVA24v strains from this study when compared to the close strains. Conjunctivitis patients with one of the three symptoms (weakness, fever and blurred vision) had a higher detection rate of HAdV. Eleven known HAdV genotypes (HAdV-B3, -B14, -B21, HAdV-D8, -D37, -D42, -D53, -D64, -D85, -D115 and HAdV-E4) and 7 unknown genotypes were detected in 2020-2024, with HAdV-D37 (30.9 %), HAdV-D115 (21.1 %) and HAdV-B3 (17.1 %) being the three predominant genotypes.</p><p><strong>Conclusions: </strong>The unique genetic characteristics were observed in Shenzhen CVA24v strains. HAdVs associated with conjunctivitis exhibited a high degree of genotypic diversity in Shenzhen, and HAdV-D115 related to conjunctivitis was first reported in this study.</p>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"180 ","pages":"105852"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144812096","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01DOI: 10.1016/j.jcv.2025.105875
Huimin Liu , Wenting Chen , Yongjie Liang , Jing Wang , Lijian Ran , Shilian Li , Yi Wu , Zixuan He , Xuemei Kuang , Jie Xia , Li Jiang , Xuqing Zhang , Qing Mao
Background and aims
This study aimed to evaluate the efficacy of expanding antiviral indications in preventing mother-to-child transmission (MTCT) by analyzing hepatitis B virus (HBV) markers in neonatal umbilical-cord-blood (UCB) samples.
Methods
We conducted a real-world study of pregnant women aged >30 years with chronic hepatitis B (CHB) who received antenatal care between January 2022 and June 2024. Maternal HBV markers at 24–28 weeks of gestation and serum biochemical parameters at delivery were obtained. For infants, HBV markers were measured in UCB at birth and in venous blood at 7–12 months of age. Logistic regression analysis was used to identify the maternal factors associated with UCB outcomes.
Results
A total of 171 pregnant women with CHB were included. Antiviral therapy had been started before conception in 31.0 % (53/171). At 24–28 weeks of gestation, 87.8 % (150/171) had HBV DNA <5.3 Log10 IU/mL, 56.1 % (96/171) had hepatitis B surface antigen (HBsAg) >3000 IU/mL and 32.7 % (56/171) were hepatitis B e antigen (HBeAg)-positive. At delivery, HBeAg-negative mothers had a median age of 33.4 ± 3.1 years, compared with 32.5 ± 2.9 years for those who were HBeAg-positive, respectively (t = 1.667, P = 0.970). 36 of 171 (21.1 %) infants were positive for HBsAg, including 12 of 115 born to HBeAg-negative mothers and 24 of 56 born to HBeAg-positive mothers (χ2 = 23.82, P < 0.001). 41 of 171 (24.0 %) infants were positive for HBeAg, and all were born to HBeAg-positive mothers. HBV DNA and RNA were undetectable in all UCB samples, whereas hepatitis B core antibody (HBcAb) was present in every specimen. Receiver operating characteristic curve (ROC) analysis identified maternal HBeAg levels (AUC=0.683, cut-off value=0.494 COI) and maternal HBV DNA load (AUC=0.645, cut-off value=0.311 Log10 IU/mL) as predictive of UCB HBsAg-positivity. No infants were infected with HBV, as confirmed by post-vaccination serologic testing (PVST).
Conclusions
Following the expansion of antiviral therapy indications, administering therapy to pregnant women aged >30 years with detectable HBV DNA effectively reduced HBsAg, HBV DNA and even HBV RNA level in infants’ UCB. Maternal HBeAg status was significantly associated with both HBsAg and HBeAg positivity in UCB.
{"title":"More aggressive initiation of antiviral treatment contributes to blocking mother-to-child transmission of HBV DNA & RNA in neonatal umbilical cord blood","authors":"Huimin Liu , Wenting Chen , Yongjie Liang , Jing Wang , Lijian Ran , Shilian Li , Yi Wu , Zixuan He , Xuemei Kuang , Jie Xia , Li Jiang , Xuqing Zhang , Qing Mao","doi":"10.1016/j.jcv.2025.105875","DOIUrl":"10.1016/j.jcv.2025.105875","url":null,"abstract":"<div><h3>Background and aims</h3><div>This study aimed to evaluate the efficacy of expanding antiviral indications in preventing mother-to-child transmission (MTCT) by analyzing hepatitis B virus (HBV) markers in neonatal umbilical-cord-blood (UCB) samples.</div></div><div><h3>Methods</h3><div>We conducted a real-world study of pregnant women aged >30 years with chronic hepatitis B (CHB) who received antenatal care between January 2022 and June 2024. Maternal HBV markers at 24–28 weeks of gestation and serum biochemical parameters at delivery were obtained. For infants, HBV markers were measured in UCB at birth and in venous blood at 7–12 months of age. Logistic regression analysis was used to identify the maternal factors associated with UCB outcomes.</div></div><div><h3>Results</h3><div>A total of 171 pregnant women with CHB were included. Antiviral therapy had been started before conception in 31.0 % (53/171). At 24–28 weeks of gestation, 87.8 % (150/171) had HBV DNA <5.3 Log<sub>10</sub> IU/mL, 56.1 % (96/171) had hepatitis B surface antigen (HBsAg) >3000 IU/mL and 32.7 % (56/171) were hepatitis B e antigen (HBeAg)-positive. At delivery, HBeAg-negative mothers had a median age of 33.4 ± 3.1 years, compared with 32.5 ± 2.9 years for those who were HBeAg-positive, respectively (t = 1.667, <em>P</em> = 0.970). 36 of 171 (21.1 %) infants were positive for HBsAg, including 12 of 115 born to HBeAg-negative mothers and 24 of 56 born to HBeAg-positive mothers (χ<sup>2</sup> = 23.82, <em>P</em> < 0.001). 41 of 171 (24.0 %) infants were positive for HBeAg, and all were born to HBeAg-positive mothers. HBV DNA and RNA were undetectable in all UCB samples, whereas hepatitis B core antibody (HBcAb) was present in every specimen. Receiver operating characteristic curve (ROC) analysis identified maternal HBeAg levels (AUC=0.683, cut-off value=0.494 COI) and maternal HBV DNA load (AUC=0.645, cut-off value=0.311 Log<sub>10</sub> IU/mL) as predictive of UCB HBsAg-positivity. No infants were infected with HBV, as confirmed by post-vaccination serologic testing (PVST).</div></div><div><h3>Conclusions</h3><div>Following the expansion of antiviral therapy indications, administering therapy to pregnant women aged >30 years with detectable HBV DNA effectively reduced HBsAg, HBV DNA and even HBV RNA level in infants’ UCB. Maternal HBeAg status was significantly associated with both HBsAg and HBeAg positivity in UCB.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"181 ","pages":"Article 105875"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145268018","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01DOI: 10.1016/j.jcv.2025.105876
Rachel Jackson , Adinah Marks , William L. Irving , Kathryn Jack
Background
Hepatitis B virus (HBV) infection is an important cause of liver disease-related mortality and morbidity. The World Health Organisation aims to eliminate this as a public health concern by 2030 and as such the key international guidelines recommend that all patients are reviewed regularly to observe for preventable signs of disease progression. This requires life-long engagement with specialist services, but some patients fall out of the care pathway and become lost to follow-up.
Objectives
This study sought to identify and re-engage patients with HBV who were lost to follow up (LTFU), defined as any HBsAg positive patient who had not been seen in the hepatology outpatient service since 31st December 2021, excluding those with acute infection.
Study design
A retrospective case finding and re-engagement healthcare service improvement exercise was conducted to identify and contact individuals with HBV diagnosed between June 2007 and the end of December 2021 who were lost-to-follow-up.
Results
One third of the HBsAg positive cohort were lost to follow-up (32.9 %, n = 506/1539). Of this group, 145 people were still living in the hospital’s catchment area, yet only 60 people could be contacted by telephone of whom 50 returned to clinic. More than 12 % of patients were HBeAg positive at their last clinic visit, and almost one quarter (23.2 %) had an abnormally raised ALT. There was extensive ethnic heterogeneity with 65 languages spoken among 474 people. We successfully re-engaged 10.07 % (51/506) back into care.
Conclusions
Patients with potentially progressive HBV-related liver disease are falling out of the care pathway with the attendant long-term problems that failure to control their infection may have.
{"title":"Identifying people with chronic hepatitis B virus who are lost to clinical follow up: A retrospective case finding and re-engagement service improvement exercise","authors":"Rachel Jackson , Adinah Marks , William L. Irving , Kathryn Jack","doi":"10.1016/j.jcv.2025.105876","DOIUrl":"10.1016/j.jcv.2025.105876","url":null,"abstract":"<div><h3>Background</h3><div>Hepatitis B virus (HBV) infection is an important cause of liver disease-related mortality and morbidity. The World Health Organisation aims to eliminate this as a public health concern by 2030 and as such the key international guidelines recommend that all patients are reviewed regularly to observe for preventable signs of disease progression. This requires life-long engagement with specialist services, but some patients fall out of the care pathway and become lost to follow-up.</div></div><div><h3>Objectives</h3><div>This study sought to identify and re-engage patients with HBV who were lost to follow up (LTFU), defined as any HBsAg positive patient who had not been seen in the hepatology outpatient service since 31st December 2021, excluding those with acute infection.</div></div><div><h3>Study design</h3><div>A retrospective case finding and re-engagement healthcare service improvement exercise was conducted to identify and contact individuals with HBV diagnosed between June 2007 and the end of December 2021 who were lost-to-follow-up.</div></div><div><h3>Results</h3><div>One third of the HBsAg positive cohort were lost to follow-up (32.9 %, n = 506/1539). Of this group, 145 people were still living in the hospital’s catchment area, yet only 60 people could be contacted by telephone of whom 50 returned to clinic. More than 12 % of patients were HBeAg positive at their last clinic visit, and almost one quarter (23.2 %) had an abnormally raised ALT. There was extensive ethnic heterogeneity with 65 languages spoken among 474 people. We successfully re-engaged 10.07 % (51/506) back into care.</div></div><div><h3>Conclusions</h3><div>Patients with potentially progressive HBV-related liver disease are falling out of the care pathway with the attendant long-term problems that failure to control their infection may have.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"181 ","pages":"Article 105876"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145220248","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-07-22DOI: 10.1016/j.jcv.2025.105843
Manfred Accrombessi, Cheryl Johnson, Alaleh Abadpour, Jean De Dieu Anoubissi, Joseph Fokam, Araz Chiloyan, Iryna Andrianova, Hamakwa Mantina, Agai Kherbino Akec, Fatou Ousmane Sall, Abdelaye Keita, Elizabeth Telan, Adoum Fouda Abderrazzack, Chatté Adawaye, Rose Wafula, Stephen Ayisi-Addo, Monkoe S Leqheka, Jacob Lusekelo Mwambeta, Jeremie Muwonga Masidi, Dramane Kania, Anita Sands, Rachel Baggaley, Jean-François Etard, Céline Lastrucci
This study highlights the importance of verifying HIV testing algorithms to reduce the risk of misdiagnoses caused by common false reactivity. Between 2020 and 2023, WHO supported 14 countries to assess rates of false reactivity and shared false reactivity across HIV rapid diagnostic tests (RDTs) used in HIV testing services. The study involved 26,278 results from 22 different RDT products, with sample sizes ranging from 100 to 302 results per country. The number of RDT products assessed varied between 4 and 13 per country. False reactivity rates ranged from 0 % to 3.32 %, with one country reporting a high false reactivity rate of over 4 % for one RDT. Five countries have no shared false reactivity between RDTs, while the remaining eight countries shared false reactivity across one to six pairs of RDT products. These findings were used to inform national policy, with more than 90 % of countries introducing new RDT products into their HIV testing algorithm based on these results. The study concludes that rates of false reactivity and shared false reactivity between RDT products vary across countries. Therefore, conducting verification studies is crucial for updating national HIV testing algorithms and ensuring accurate diagnosis while also facilitating the market entry of new HIV testing products.
{"title":"Heterogeneity of false reactivity profiles of HIV assays while optimizing national HIV testing algorithms: Findings from a multi-country analysis.","authors":"Manfred Accrombessi, Cheryl Johnson, Alaleh Abadpour, Jean De Dieu Anoubissi, Joseph Fokam, Araz Chiloyan, Iryna Andrianova, Hamakwa Mantina, Agai Kherbino Akec, Fatou Ousmane Sall, Abdelaye Keita, Elizabeth Telan, Adoum Fouda Abderrazzack, Chatté Adawaye, Rose Wafula, Stephen Ayisi-Addo, Monkoe S Leqheka, Jacob Lusekelo Mwambeta, Jeremie Muwonga Masidi, Dramane Kania, Anita Sands, Rachel Baggaley, Jean-François Etard, Céline Lastrucci","doi":"10.1016/j.jcv.2025.105843","DOIUrl":"10.1016/j.jcv.2025.105843","url":null,"abstract":"<p><p>This study highlights the importance of verifying HIV testing algorithms to reduce the risk of misdiagnoses caused by common false reactivity. Between 2020 and 2023, WHO supported 14 countries to assess rates of false reactivity and shared false reactivity across HIV rapid diagnostic tests (RDTs) used in HIV testing services. The study involved 26,278 results from 22 different RDT products, with sample sizes ranging from 100 to 302 results per country. The number of RDT products assessed varied between 4 and 13 per country. False reactivity rates ranged from 0 % to 3.32 %, with one country reporting a high false reactivity rate of over 4 % for one RDT. Five countries have no shared false reactivity between RDTs, while the remaining eight countries shared false reactivity across one to six pairs of RDT products. These findings were used to inform national policy, with more than 90 % of countries introducing new RDT products into their HIV testing algorithm based on these results. The study concludes that rates of false reactivity and shared false reactivity between RDT products vary across countries. Therefore, conducting verification studies is crucial for updating national HIV testing algorithms and ensuring accurate diagnosis while also facilitating the market entry of new HIV testing products.</p>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"180 ","pages":"105843"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12439682/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144816831","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-08-06DOI: 10.1016/j.jcv.2025.105853
Anja Oštrbenk, Helle Pedersen, Mario Poljak, Jesper Bonde
A key parameter for the continued success of cervical cancer screening is quality-controlled use of human papillomavirus (HPV) tests that are clinically validated according to international guidelines. The clinical accuracy for cervical screening of the Allplex HPV HR Detection (Allplex), which concurrently detects and distinguishes 12 high-risk HPV types (16,18,31,33,35,39,45,51,52,56,58,59) and HPV66 and HPV68 was assessed on SurePath samples by comparing its performance to the second-generation comparator BD Onclarity HPV assay (Onclarity). The absolute clinical sensitivity, assessed on 76 samples derived from a screening population with underlying CIN2+, of Allplex and Onclarity was 98.7 % (95 % CI, 92.9-100.0 %) and 100.0 % (95 % CI, 95.2-100.0 %), respectively, with relative sensitivity of Allplex of 0.99 (95 % CI; 0.96-1.01). The absolute clinical specificity, assessed on 801 consecutive clinician-collected cervical samples obtained from women 30 to 59 years old attending the routine Danish cervical screening program, for Allplex and Onclarity was 92.5 % (95 % CI, 90.4-94.2 %) and 92.5 % (95 % CI, 90.4-94.2 %), respectively, with relative specificity of Allplex of 1.00 (95 % CI; 0.99-1.01). With thresholds mandated by international guidelines of ≥90 % for relative clinical sensitivity (p = 0.001) and ≥98 % for relative clinical specificity (p = 0.0018), Allplex was non-inferior to Onclarity. Excellent intra- and inter-laboratory agreement of Allplex was observed, both overall (99.2 % and 99.6 %) and at the genotype level (range: 99.6-100.0 %). By fulfilling all guideline requirements for clinical sensitivity, specificity, and reproducibility, Allplex can be considered clinically validated for primary cervical screening using clinician-collected SurePath samples. With this study, Allplex also meets the criteria for a second-generation HPV comparator test.
{"title":"The Allplex HPV HR Detection assay fulfils international guideline requirements for primary cervical screening on SurePath samples and qualifies as a second-generation HPV comparator test.","authors":"Anja Oštrbenk, Helle Pedersen, Mario Poljak, Jesper Bonde","doi":"10.1016/j.jcv.2025.105853","DOIUrl":"10.1016/j.jcv.2025.105853","url":null,"abstract":"<p><p>A key parameter for the continued success of cervical cancer screening is quality-controlled use of human papillomavirus (HPV) tests that are clinically validated according to international guidelines. The clinical accuracy for cervical screening of the Allplex HPV HR Detection (Allplex), which concurrently detects and distinguishes 12 high-risk HPV types (16,18,31,33,35,39,45,51,52,56,58,59) and HPV66 and HPV68 was assessed on SurePath samples by comparing its performance to the second-generation comparator BD Onclarity HPV assay (Onclarity). The absolute clinical sensitivity, assessed on 76 samples derived from a screening population with underlying CIN2+, of Allplex and Onclarity was 98.7 % (95 % CI, 92.9-100.0 %) and 100.0 % (95 % CI, 95.2-100.0 %), respectively, with relative sensitivity of Allplex of 0.99 (95 % CI; 0.96-1.01). The absolute clinical specificity, assessed on 801 consecutive clinician-collected cervical samples obtained from women 30 to 59 years old attending the routine Danish cervical screening program, for Allplex and Onclarity was 92.5 % (95 % CI, 90.4-94.2 %) and 92.5 % (95 % CI, 90.4-94.2 %), respectively, with relative specificity of Allplex of 1.00 (95 % CI; 0.99-1.01). With thresholds mandated by international guidelines of ≥90 % for relative clinical sensitivity (p = 0.001) and ≥98 % for relative clinical specificity (p = 0.0018), Allplex was non-inferior to Onclarity. Excellent intra- and inter-laboratory agreement of Allplex was observed, both overall (99.2 % and 99.6 %) and at the genotype level (range: 99.6-100.0 %). By fulfilling all guideline requirements for clinical sensitivity, specificity, and reproducibility, Allplex can be considered clinically validated for primary cervical screening using clinician-collected SurePath samples. With this study, Allplex also meets the criteria for a second-generation HPV comparator test.</p>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"180 ","pages":"105853"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804233","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-08-05DOI: 10.1016/j.jcv.2025.105849
Sébastien Lhomme, Isabelle Da Silva, Oriane Cabanel, Estelle Raguin, Guillaume Martin-Blondel, Nassim Kamar, Jacques Izopet, Florence Abravanel
Introduction: We evaluated the performance of the automated AltoStar JCV PCR platform for detecting and quantifying JC Polyomavirus (JCPyV) DNA in samples including urine, plasma and cerebrospinal fluid.
Methods and results: Using the NIBSC JCPyV DNA standard, the assay was linear from 1 to 6 log IU/mL standard and the limit of detection determined by Probit analysis corresponded to 9.3 [95 % CI: 7.0-16.5] IU/mL. Specificity was accessed by testing urines containing a high concentration of BK Polyomavirus; none tested positive for JCPyV. The intra-run and inter-run standard deviations were 0.02 IU/mL and 0.35 IU/mL, respectively. Lastly, clinical performance was determined by testing 45 samples quantified previously with a laboratory-developed test (LDT). The assays were concordant for 42/45 samples. One of the 3 samples that tested negative with the AltoStar assay had a low JCpyV DNA concentration. We were unable to re-test the 3 negative samples due to insufficient volume. A conservation problem could not be ruled out for the 3 samples with discordant results.
Conclusions: The AltoStar platform enables highly accurate testing for the detection and quantification of JCPyV DNA with very low limit of detection. This allowed us to shorten turnaround times and save time for technical staff.
{"title":"Performance of a commercial assay for detecting JC Polyomavirus DNA in human samples.","authors":"Sébastien Lhomme, Isabelle Da Silva, Oriane Cabanel, Estelle Raguin, Guillaume Martin-Blondel, Nassim Kamar, Jacques Izopet, Florence Abravanel","doi":"10.1016/j.jcv.2025.105849","DOIUrl":"10.1016/j.jcv.2025.105849","url":null,"abstract":"<p><strong>Introduction: </strong>We evaluated the performance of the automated AltoStar JCV PCR platform for detecting and quantifying JC Polyomavirus (JCPyV) DNA in samples including urine, plasma and cerebrospinal fluid.</p><p><strong>Methods and results: </strong>Using the NIBSC JCPyV DNA standard, the assay was linear from 1 to 6 log IU/mL standard and the limit of detection determined by Probit analysis corresponded to 9.3 [95 % CI: 7.0-16.5] IU/mL. Specificity was accessed by testing urines containing a high concentration of BK Polyomavirus; none tested positive for JCPyV. The intra-run and inter-run standard deviations were 0.02 IU/mL and 0.35 IU/mL, respectively. Lastly, clinical performance was determined by testing 45 samples quantified previously with a laboratory-developed test (LDT). The assays were concordant for 42/45 samples. One of the 3 samples that tested negative with the AltoStar assay had a low JCpyV DNA concentration. We were unable to re-test the 3 negative samples due to insufficient volume. A conservation problem could not be ruled out for the 3 samples with discordant results.</p><p><strong>Conclusions: </strong>The AltoStar platform enables highly accurate testing for the detection and quantification of JCPyV DNA with very low limit of detection. This allowed us to shorten turnaround times and save time for technical staff.</p>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"180 ","pages":"105849"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144804232","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-01Epub Date: 2025-08-06DOI: 10.1016/j.jcv.2025.105850
Romina Salpini, Lorenzo Piermatteo, Gian Paolo Caviglia, Ada Bertoli, Maurizia Rossana Brunetto, Bianca Bruzzone, Annapaola Callegaro, Cinzia Caudai, Daniela Cavallone, Luchino Chessa, Fernando Coghe, Nicola Coppola, Nunzia Cuomo, Stefano D'Anna, Mariantonietta Di Stefano, Floriana Facchetti, Claudio Farina, Donatella Ferraro, Elisa Franchin, Daniela Francisci, Silvia Galli, Anna Rosa Garbuglia, William Gennari, Valeria Ghisetti, Pietro Lampertico, Sergio Lo Caputo, Nadia Marascio, Stefano Menzo, Valeria Micheli, Grazia Anna Niro, Antonella Olivero, Pierpaolo Paba, Concetta Ilenia Palermo, Orazio Palmieri, Stefania Paolucci, Mariantonietta Pisaturo, Teresa Pollicino, Giuseppina Raffa, Teresa Santantonio, Giulia Torre, Ombretta Turriziani, Sergio Uzzau, Sara Colonia Uceda Renteria, Marialinda Vatteroni, Maurizio Zazzi, Antonio Craxì, Francesca Ceccherini-Silberstein, Valentina Svicher, Marco Arosio, Sabrina Bastianelli, Annamaria Gentile, Federica A M Giardina, Anna Gidari, Rosalba Govoni, Gabriele Ibba, Alessandro Loglio, Alessandra Lombardi, Chiara Mascarella, Fabrizio Maggi, Giovanni Matera, Chiara Mazzei, Maria Grazia Milia, Angela Quirino, Adriana Raddi, Rosetta Scioscia, Sara Tagliazucchi, Michele Totaro, Rea Valaperta
Introduction: A reliable quantification of hepatitis D virus (HDV) RNA is of paramount importance for monitoring patients under antiviral therapy. This quality control study compares the diagnostic performances of quantitative HDV-RNA assays used in clinical practice.
Methods: Two HDV-RNA sample panels were quantified in 30 centers by RoboGene (N = 9 laboratories), EurobioPlex (N = 7), RealStar (N = 4), AltoStar (N = 1), Bosphore (N = 3), Bosphore-on-InGenius (N = 1), Dia.Pro (N = 2), Nuclear-Laser-Medicine (N = 1) and 3 in-house assays. Panel A and B comprised 8 serial dilutions of WHO/HDV standard (range: 0.5-5.0 log10 IU/ml) and 20 clinical samples (range: 0.5-6.0 log10 IU/ml), respectively. The following parameters were determined: sensitivity by 95 % LOD (limit of detection), precision by intra- and inter-run CV (coefficient of variation), accuracy by the differences between expected-observed HDV-RNA, linearity by linear regression analysis.
Results: 95 % LOD varied across assays and centers underlining heterogeneous sensitivities: AltoStar had the lowest 95 % LOD (3 IU/ml) followed by RealStar (10 [min-max: 3-316] IU/ml), Bosphore-on-InGenius (10 IU/ml), RoboGene (31 [3-316] IU/ml), Nuclear-Laser-Medicine (31 IU/ml) and EuroBioplex (100 [100-316] IU/ml). Moreover, 6 assays (RoboGene, EurobioPlex, RealStar, AltoStar, Nuclear-Laser-Medicine and In-house) showed <0.5 log10 IU/ml differences between expected and observed HDV-RNA for all dilutions while other assays had >1 log10 IU/ml underestimations. RealStar, Bosphore-on-InGenius and EurobioPlex had the highest precision (mean intra-run CV < 20 %). Inter-run CV was higher for all assays, with CVs < 25 % for RealStar, AltoStar, Nuclear-Laser-Medicine and EurobioPlex. Seven assays (RoboGene/AltoStar/RealStar/EurobioPlex/Nuclear-Laser-Medicine/In-house) showed a good linearity (R2 > 0.90), but for HDV-RNA < 1000 IU/ml only Bosphore-on-InGenius, AltoStar, RealStar and Robogene showed a R2 > 0.85.
Conclusions: This study underlines heterogeneous sensitivities (inter- and intraassays), that could hamper proper HDV-RNA quantification, particularly at low viral loads. This raises the need to improve the diagnostic performance of most assays for properly identifying virological response to anti-HDV drugs.
{"title":"Comparison of diagnostic performances of HDV-RNA quantification assays used in clinical practice: Results from a national quality control multicenter study.","authors":"Romina Salpini, Lorenzo Piermatteo, Gian Paolo Caviglia, Ada Bertoli, Maurizia Rossana Brunetto, Bianca Bruzzone, Annapaola Callegaro, Cinzia Caudai, Daniela Cavallone, Luchino Chessa, Fernando Coghe, Nicola Coppola, Nunzia Cuomo, Stefano D'Anna, Mariantonietta Di Stefano, Floriana Facchetti, Claudio Farina, Donatella Ferraro, Elisa Franchin, Daniela Francisci, Silvia Galli, Anna Rosa Garbuglia, William Gennari, Valeria Ghisetti, Pietro Lampertico, Sergio Lo Caputo, Nadia Marascio, Stefano Menzo, Valeria Micheli, Grazia Anna Niro, Antonella Olivero, Pierpaolo Paba, Concetta Ilenia Palermo, Orazio Palmieri, Stefania Paolucci, Mariantonietta Pisaturo, Teresa Pollicino, Giuseppina Raffa, Teresa Santantonio, Giulia Torre, Ombretta Turriziani, Sergio Uzzau, Sara Colonia Uceda Renteria, Marialinda Vatteroni, Maurizio Zazzi, Antonio Craxì, Francesca Ceccherini-Silberstein, Valentina Svicher, Marco Arosio, Sabrina Bastianelli, Annamaria Gentile, Federica A M Giardina, Anna Gidari, Rosalba Govoni, Gabriele Ibba, Alessandro Loglio, Alessandra Lombardi, Chiara Mascarella, Fabrizio Maggi, Giovanni Matera, Chiara Mazzei, Maria Grazia Milia, Angela Quirino, Adriana Raddi, Rosetta Scioscia, Sara Tagliazucchi, Michele Totaro, Rea Valaperta","doi":"10.1016/j.jcv.2025.105850","DOIUrl":"10.1016/j.jcv.2025.105850","url":null,"abstract":"<p><strong>Introduction: </strong>A reliable quantification of hepatitis D virus (HDV) RNA is of paramount importance for monitoring patients under antiviral therapy. This quality control study compares the diagnostic performances of quantitative HDV-RNA assays used in clinical practice.</p><p><strong>Methods: </strong>Two HDV-RNA sample panels were quantified in 30 centers by RoboGene (N = 9 laboratories), EurobioPlex (N = 7), RealStar (N = 4), AltoStar (N = 1), Bosphore (N = 3), Bosphore-on-InGenius (N = 1), Dia.Pro (N = 2), Nuclear-Laser-Medicine (N = 1) and 3 in-house assays. Panel A and B comprised 8 serial dilutions of WHO/HDV standard (range: 0.5-5.0 log10 IU/ml) and 20 clinical samples (range: 0.5-6.0 log10 IU/ml), respectively. The following parameters were determined: sensitivity by 95 % LOD (limit of detection), precision by intra- and inter-run CV (coefficient of variation), accuracy by the differences between expected-observed HDV-RNA, linearity by linear regression analysis.</p><p><strong>Results: </strong>95 % LOD varied across assays and centers underlining heterogeneous sensitivities: AltoStar had the lowest 95 % LOD (3 IU/ml) followed by RealStar (10 [min-max: 3-316] IU/ml), Bosphore-on-InGenius (10 IU/ml), RoboGene (31 [3-316] IU/ml), Nuclear-Laser-Medicine (31 IU/ml) and EuroBioplex (100 [100-316] IU/ml). Moreover, 6 assays (RoboGene, EurobioPlex, RealStar, AltoStar, Nuclear-Laser-Medicine and In-house) showed <0.5 log10 IU/ml differences between expected and observed HDV-RNA for all dilutions while other assays had >1 log10 IU/ml underestimations. RealStar, Bosphore-on-InGenius and EurobioPlex had the highest precision (mean intra-run CV < 20 %). Inter-run CV was higher for all assays, with CVs < 25 % for RealStar, AltoStar, Nuclear-Laser-Medicine and EurobioPlex. Seven assays (RoboGene/AltoStar/RealStar/EurobioPlex/Nuclear-Laser-Medicine/In-house) showed a good linearity (R<sup>2</sup> > 0.90), but for HDV-RNA < 1000 IU/ml only Bosphore-on-InGenius, AltoStar, RealStar and Robogene showed a R<sup>2</sup> > 0.85.</p><p><strong>Conclusions: </strong>This study underlines heterogeneous sensitivities (inter- and intraassays), that could hamper proper HDV-RNA quantification, particularly at low viral loads. This raises the need to improve the diagnostic performance of most assays for properly identifying virological response to anti-HDV drugs.</p>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"180 ","pages":"105850"},"PeriodicalIF":3.4,"publicationDate":"2025-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144816830","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-25DOI: 10.1016/j.jcv.2025.105874
Bo Shu , William G. Davis , Ji Liu , Beth K. Thielen , Sarah Bistodeau , Brian Lynch , Christine M. Warnes , Jimma Liddell , Anna K. Strain , Jaime Christensen , Phili Wong , Natasha Burnett , Todd C. Davis , Marie K. Kirby
Influenza C virus (ICV) usually causes a mild upper respiratory tract infection in children and those infected are frequently co-infected with other respiratory viruses. However, there have only been a few hundred documented cases of ICV infection in humans as of the end of 2024. To better understand the epidemiology and clinical impact of ICVs, we developed an influenza C real-time RT-PCR (InfC rRT-PCR) assay that targets a highly conserved region of the matrix gene segment of ICVs. The analytical sensitivity evaluation demonstrated that the InfC rRT-PCR assay was highly sensitive, as it was able to detect as few as five RNA copies per PCR reaction and had robust reactivity over a range of viral RNAs from historical and recent ICVs. The analytical specificity evaluation confirmed the assay did not cross-react with any influenza A or B viruses tested, including several animal-origin viruses, or other common non-influenza respiratory viruses. The performance evaluation on clinical specimens demonstrated the assay was highly sensitive and specific for the detection of ICVs.
{"title":"Design and performance of a real-time RT-PCR assay for detection of influenza C viruses","authors":"Bo Shu , William G. Davis , Ji Liu , Beth K. Thielen , Sarah Bistodeau , Brian Lynch , Christine M. Warnes , Jimma Liddell , Anna K. Strain , Jaime Christensen , Phili Wong , Natasha Burnett , Todd C. Davis , Marie K. Kirby","doi":"10.1016/j.jcv.2025.105874","DOIUrl":"10.1016/j.jcv.2025.105874","url":null,"abstract":"<div><div>Influenza C virus (ICV) usually causes a mild upper respiratory tract infection in children and those infected are frequently co-infected with other respiratory viruses. However, there have only been a few hundred documented cases of ICV infection in humans as of the end of 2024. To better understand the epidemiology and clinical impact of ICVs, we developed an influenza C real-time RT-PCR (InfC rRT-PCR) assay that targets a highly conserved region of the matrix gene segment of ICVs. The analytical sensitivity evaluation demonstrated that the InfC rRT-PCR assay was highly sensitive, as it was able to detect as few as five RNA copies per PCR reaction and had robust reactivity over a range of viral RNAs from historical and recent ICVs. The analytical specificity evaluation confirmed the assay did not cross-react with any influenza A or B viruses tested, including several animal-origin viruses, or other common non-influenza respiratory viruses. The performance evaluation on clinical specimens demonstrated the assay was highly sensitive and specific for the detection of ICVs.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"181 ","pages":"Article 105874"},"PeriodicalIF":3.4,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145206489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-22DOI: 10.1016/j.jcv.2025.105873
Martin Wälti , Marco Natali , Adrian Sanden , Remo Leisi , Svenja Rohde , Rick Alexander , Jonas Sieber , Christoph Zürcher , Björn Keiner , Kelley Hyatt , Eva Quinley , Joyce Castaneda , Michael Schiffer , Denis Klochkov , Eleonora Widmer
Background
Parvovirus B19 (B19V) infection is characterized by measurable bloodstream biomarkers, which vary over time indicating different phases of infection. High B19V DNA levels in the bloodstream are associated with infectivity, impacting the eligibility of source plasma donors.
Objectives
Characterize temporal changes in B19V epidemiological rates from January 2018 to April 2025 and to investigate the longitudinal dynamics of the diagnostic biomarkers and infectivity in subclinical infections among US source plasma donors.
Results
B19V positivity rates in CSL’s US source plasma donors decreased to a very low level during the COVID-19 pandemic and until Spring 2023 and re-emerged in two waves with peak rates ≥ 2.2-fold higher compared to pre-COVID data (2018 – 2020). During the exponential infection phase, B19V DNA doubled every 3–5 h, leading to high titers up to 10^12 IU/mL and peak infectivity of ∼10^8 TCID50/mL. Infectivity ceased when anti-B19V IgG reached a plateau or 14 days after DNA titer dropped below 4.8 × 10^5 IU/mL.
Conclusions
The re-emergence of B19V with high positivity rates highlights the relevance of donation testing for plasma. During the month from infection onset, B19V DNAemia correlates well with the measured infectious titer. B19V infectivity in plasma disappears within a month while DNA can persist in the plasma in absence of detectable infectivity, likely due to the immune response as characterized by the immunological markers.
{"title":"Assessing B19V epidemiology and viremia dynamics in plasma donors: A comprehensive analysis","authors":"Martin Wälti , Marco Natali , Adrian Sanden , Remo Leisi , Svenja Rohde , Rick Alexander , Jonas Sieber , Christoph Zürcher , Björn Keiner , Kelley Hyatt , Eva Quinley , Joyce Castaneda , Michael Schiffer , Denis Klochkov , Eleonora Widmer","doi":"10.1016/j.jcv.2025.105873","DOIUrl":"10.1016/j.jcv.2025.105873","url":null,"abstract":"<div><h3>Background</h3><div>Parvovirus B19 (B19V) infection is characterized by measurable bloodstream biomarkers, which vary over time indicating different phases of infection. High B19V DNA levels in the bloodstream are associated with infectivity, impacting the eligibility of source plasma donors.</div></div><div><h3>Objectives</h3><div>Characterize temporal changes in B19V epidemiological rates from January 2018 to April 2025 and to investigate the longitudinal dynamics of the diagnostic biomarkers and infectivity in subclinical infections among US source plasma donors.</div></div><div><h3>Results</h3><div>B19V positivity rates in CSL’s US source plasma donors decreased to a very low level during the COVID-19 pandemic and until Spring 2023 and re-emerged in two waves with peak rates ≥ 2.2-fold higher compared to pre-COVID data (2018 – 2020). During the exponential infection phase, B19V DNA doubled every 3–5 h, leading to high titers up to 10^12 IU/mL and peak infectivity of ∼10^8 TCID<sub>50</sub>/mL. Infectivity ceased when anti-B19V IgG reached a plateau or 14 days after DNA titer dropped below 4.8 × 10^5 IU/mL.</div></div><div><h3>Conclusions</h3><div>The re-emergence of B19V with high positivity rates highlights the relevance of donation testing for plasma. During the month from infection onset, B19V DNAemia correlates well with the measured infectious titer. B19V infectivity in plasma disappears within a month while DNA can persist in the plasma in absence of detectable infectivity, likely due to the immune response as characterized by the immunological markers.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"181 ","pages":"Article 105873"},"PeriodicalIF":3.4,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145292347","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-19DOI: 10.1016/j.jcv.2025.105872
Carlo Fischer , Anges Yadouleton , Miguel Mauricio Cabada , Miladi Gatty Nogueira , Marta Piche-Ovares , Stephane Sohou , César Augusto Cabezas Sánchez , Patricia T. Bozza , María Paquita García Mendoza , Eduardo Gotuzzo , Fernando Augusto Bozza , Jan Felix Drexler
Background
Knowledge of epidemiology, pathogenesis, and public health burden is scarce for many arthropod-borne viruses (arboviruses). Insufficient knowledge is partly attributable to the lack of exhaustive laboratory diagnostics due to resource limitations. Among arboviruses, arthritogenic and encephalitogenic alphaviruses are globally widespread, can cause severe disease, and can co-occur regionally.
Objectives
We developed and validated a multiplexed real-time reverse transcription-PCR assay for the detection of all alphaviruses commonly causing human disease except Barmah Forest virus.
Study design
The assay combines five antigenic complex-specific assays and one Chikungunya virus (CHIKV)-specific assay in a single parallelized reaction.
Results
Comparisons with previously published PCR-based protocols for broad alphavirus detection using 20 different human-pathogenic alphaviruses revealed a significantly higher sensitivity of the new multiplexed assay (Fisher’s exact test, p < 0.0001). Detection limits with the new assay ranged from 0.83 cps/μl of extracted O’nyong-nyong virus to 33.05 cps/μl of extracted Western equine encephalitis virus. Antigenic complexes could be clearly differentiated by reactivity, Ct values (t-test, p < 0.0025) and signal intensities (t-test, p < 0.0001), even when testing high alphavirus concentrations potentially capable of causing false-positive PCR results. Testing of high-titred cell culture supernatants of eight important non-alphaviral arboviruses, of 4308 serum samples collected from febrile patients in Benin and Peru, of seven CHIKV-positive diagnostic samples from Brazil, and of non-targeted alphaviruses confirmed excellent diagnostic performance by the new assay, including improved detection of CHIKV, Mayaro and Venezuelan equine encephalitis virus in clinical specimens.
Conclusions
Short turn-around time, applicability in resource-limited settings, antigenic complex determination, and higher sensitivity compared to previously available tests make the new assay a useful tool for alphavirus surveillance and routine patient diagnostics.
{"title":"Syndromic approach for rapid detection and differentiation of human pathogenic alphaviruses","authors":"Carlo Fischer , Anges Yadouleton , Miguel Mauricio Cabada , Miladi Gatty Nogueira , Marta Piche-Ovares , Stephane Sohou , César Augusto Cabezas Sánchez , Patricia T. Bozza , María Paquita García Mendoza , Eduardo Gotuzzo , Fernando Augusto Bozza , Jan Felix Drexler","doi":"10.1016/j.jcv.2025.105872","DOIUrl":"10.1016/j.jcv.2025.105872","url":null,"abstract":"<div><h3>Background</h3><div>Knowledge of epidemiology, pathogenesis, and public health burden is scarce for many arthropod-borne viruses (arboviruses). Insufficient knowledge is partly attributable to the lack of exhaustive laboratory diagnostics due to resource limitations. Among arboviruses, arthritogenic and encephalitogenic alphaviruses are globally widespread, can cause severe disease, and can co-occur regionally.</div></div><div><h3>Objectives</h3><div>We developed and validated a multiplexed real-time reverse transcription-PCR assay for the detection of all alphaviruses commonly causing human disease except Barmah Forest virus.</div></div><div><h3>Study design</h3><div>The assay combines five antigenic complex-specific assays and one Chikungunya virus (CHIKV)-specific assay in a single parallelized reaction.</div></div><div><h3>Results</h3><div>Comparisons with previously published PCR-based protocols for broad alphavirus detection using 20 different human-pathogenic alphaviruses revealed a significantly higher sensitivity of the new multiplexed assay (Fisher’s exact test, p < 0.0001). Detection limits with the new assay ranged from 0.83 cps/μl of extracted O’nyong-nyong virus to 33.05 cps/μl of extracted Western equine encephalitis virus. Antigenic complexes could be clearly differentiated by reactivity, Ct values (<em>t</em>-test, p < 0.0025) and signal intensities (<em>t</em>-test, p < 0.0001), even when testing high alphavirus concentrations potentially capable of causing false-positive PCR results. Testing of high-titred cell culture supernatants of eight important non-alphaviral arboviruses, of 4308 serum samples collected from febrile patients in Benin and Peru, of seven CHIKV-positive diagnostic samples from Brazil, and of non-targeted alphaviruses confirmed excellent diagnostic performance by the new assay, including improved detection of CHIKV, Mayaro and Venezuelan equine encephalitis virus in clinical specimens.</div></div><div><h3>Conclusions</h3><div>Short turn-around time, applicability in resource-limited settings, antigenic complex determination, and higher sensitivity compared to previously available tests make the new assay a useful tool for alphavirus surveillance and routine patient diagnostics.</div></div>","PeriodicalId":15517,"journal":{"name":"Journal of Clinical Virology","volume":"181 ","pages":"Article 105872"},"PeriodicalIF":3.4,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145212764","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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