为了建立牛轭湖病毒(Bayou virus,BAYV)、纽约病毒(New York virus,NYV)、厄尼诺洛峡谷病毒(El Moro Canyon virus,ELMCV)以及岛景病毒(Isla Vista virus,ISLAV)四种病毒快速筛查、诊断的核酸检测技术,本研究选用上述四种病毒的N蛋白基...为了建立牛轭湖病毒(Bayou virus,BAYV)、纽约病毒(New York virus,NYV)、厄尼诺洛峡谷病毒(El Moro Canyon virus,ELMCV)以及岛景病毒(Isla Vista virus,ISLAV)四种病毒快速筛查、诊断的核酸检测技术,本研究选用上述四种病毒的N蛋白基因作为靶标区域设计四组特异性引物探针,建立四重实时荧光定量RT-PCR检测方法。用四种病毒体外转录RNA进行灵敏度和重复性验证,并使用其他病毒细胞培养物及体外转录RNA进行特异性验证。结果显示,四重实时荧光定量PCR扩增效率均可达到95%以上,四种病毒体外转录RNA灵敏度介于1~10拷贝/μL,与单重检测方法无明显差异,且与其他病毒无交叉反应。稳定性评价结果显示,批内、批间变异系数均在3%以内。本研究所建立的检测方法具有良好的敏感性、特异性和重复性,可用于相关样本的诊断与筛查。展开更多
Background: Meningitis remains a leading cause of death among children below 5 years of age in the Democratic Republic of the Congo (DR Congo). Distinguishing children with bacterial meningitis from those with viral m...Background: Meningitis remains a leading cause of death among children below 5 years of age in the Democratic Republic of the Congo (DR Congo). Distinguishing children with bacterial meningitis from those with viral meningitis in the emergency department is sometimes difficult. Here we identified bacteria and virus in the cerebral spinal fluid (CSF) of children with meningitis. Material and Methods: This is a prospective, analytical study carried out in the Pediatrics department of Panzi Hospital in the South-Kivu province of DR Congo. Between April 2021 and March 2022, 150 of 251 collected CSF from children aged from 1 to 59 months hospitalised due to clinical meningitis at Panzi referral university hospital, Bukavu, Eastern DR Congo were sent to the Lancet laboratory for bacteria identification by a multiplex real-time PCR assay for detection of the most different viruses and bacterial species causing meningitis. Result: The used multiplex real-time PCR assay allowed us to identify germs in 24.7% of cases (37/150). We isolated bacteria in 25/37 (67.5%) cases, and viruses in 9/37 (24.3%) while virus and bacteria co-infection was detected in 3/37 (8.1%). The most frequently identified bacteria were Streptococcus pneumoniae 14/37 (37.8%) followed by Haemophilus influenzae 6/37 (16.2%). The main virus was cytomegalovirus 5/37 (3.5%). Despite the age, the most found bacterial are common in children from rural areas and unvaccinated children. Bacterial and virus co-infection were identified in 66.7% of children aged between 25 - 60 months, mainly among male children, and in all children from rural areas (100%). The overall case fatality rate was 30% and was very high among cases with co-infection CMV-Pneumococcal (66.7%), followed by Streptococcus pneumoniae (50%). Conclusion: Meningitis remains frequent among children aged from one to 59 months among Bukavu Infants. We noticed that, Children with co-infection with bacteria and viruses might need higher attention when having meningitis symptoms, as this could lead to展开更多
文摘为了建立牛轭湖病毒(Bayou virus,BAYV)、纽约病毒(New York virus,NYV)、厄尼诺洛峡谷病毒(El Moro Canyon virus,ELMCV)以及岛景病毒(Isla Vista virus,ISLAV)四种病毒快速筛查、诊断的核酸检测技术,本研究选用上述四种病毒的N蛋白基因作为靶标区域设计四组特异性引物探针,建立四重实时荧光定量RT-PCR检测方法。用四种病毒体外转录RNA进行灵敏度和重复性验证,并使用其他病毒细胞培养物及体外转录RNA进行特异性验证。结果显示,四重实时荧光定量PCR扩增效率均可达到95%以上,四种病毒体外转录RNA灵敏度介于1~10拷贝/μL,与单重检测方法无明显差异,且与其他病毒无交叉反应。稳定性评价结果显示,批内、批间变异系数均在3%以内。本研究所建立的检测方法具有良好的敏感性、特异性和重复性,可用于相关样本的诊断与筛查。
文摘Background: Meningitis remains a leading cause of death among children below 5 years of age in the Democratic Republic of the Congo (DR Congo). Distinguishing children with bacterial meningitis from those with viral meningitis in the emergency department is sometimes difficult. Here we identified bacteria and virus in the cerebral spinal fluid (CSF) of children with meningitis. Material and Methods: This is a prospective, analytical study carried out in the Pediatrics department of Panzi Hospital in the South-Kivu province of DR Congo. Between April 2021 and March 2022, 150 of 251 collected CSF from children aged from 1 to 59 months hospitalised due to clinical meningitis at Panzi referral university hospital, Bukavu, Eastern DR Congo were sent to the Lancet laboratory for bacteria identification by a multiplex real-time PCR assay for detection of the most different viruses and bacterial species causing meningitis. Result: The used multiplex real-time PCR assay allowed us to identify germs in 24.7% of cases (37/150). We isolated bacteria in 25/37 (67.5%) cases, and viruses in 9/37 (24.3%) while virus and bacteria co-infection was detected in 3/37 (8.1%). The most frequently identified bacteria were Streptococcus pneumoniae 14/37 (37.8%) followed by Haemophilus influenzae 6/37 (16.2%). The main virus was cytomegalovirus 5/37 (3.5%). Despite the age, the most found bacterial are common in children from rural areas and unvaccinated children. Bacterial and virus co-infection were identified in 66.7% of children aged between 25 - 60 months, mainly among male children, and in all children from rural areas (100%). The overall case fatality rate was 30% and was very high among cases with co-infection CMV-Pneumococcal (66.7%), followed by Streptococcus pneumoniae (50%). Conclusion: Meningitis remains frequent among children aged from one to 59 months among Bukavu Infants. We noticed that, Children with co-infection with bacteria and viruses might need higher attention when having meningitis symptoms, as this could lead to
