Development of a multiplex one-step real-time RT-PCR assay for the simultaneous detection of eight viruses associated with febrile rash illnesses

Cui, Ai‐Li; Wang, Shulei; Zhang, Qiang; Wang, Huiling; Zhu, Zhen; Li, Aqian; Song, Qinqin; Hao, Yanzhe; He, Jun; Xu, Wenbo; Zhang, Yan

doi:10.1016/j.bsheal.2020.04.003

Cited by 5 publications

(5 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…At 2 ag per reaction (equivalent to~17.3 copies/reaction), one of the replicates was found to be positive (Table 2). The PCR efficiency of the InnoPrimers-duplex qPCR was 100% for 30 bp deletion NPC genetic biomarker in this study, which represented a twofold increase in the amplicon's level after each cycle, and this efficiency value was within the acceptable range, as the acceptable range of efficiency is 90-120% [55,56].…”

Section: Analytical Sensitivity and Specificity Of The Developed Innoprimers-duplex Qpcr Assaymentioning

confidence: 50%

“…In this developed assay, r 2 values were 0.9966, which was close to 1. The PCR efficiency of the InnoPrimers-duplex qPCR was 100% for detection of 30 bp detection NPC genetic biomarker, which represented a twofold increase in the amplicon's level after each cycle, and this efficiency value was within the acceptable range, as the acceptable range of efficiency is 90-120% [55,56,82]. This developed assay is very specific for NPC disease with 100% of specificity, where the 30 bp deletion NPC genetic biomarker was not detected among all non-NPC patients and healthy individuals.…”

Section: Discussionmentioning

confidence: 89%

See 1 more Smart Citation

Design of InnoPrimers-Duplex Real-Time PCR for Detection and Treatment Response Prediction of EBV-Associated Nasopharyngeal Carcinoma Circulating Genetic Biomarker

et al. 2021

View full text Add to dashboard Cite

Nasopharyngeal carcinoma (NPC) is an epithelial tumor with high prevalence in southern China and Southeast Asia. NPC is well associated with the Epstein-Barr virus (EBV) latent membrane protein 1 (LMP1) 30 bp deletion by having its vital role in increased tumorigenicity and decreased immune recognition of EBV-related tumors. This study developed an InnoPrimers-duplex qPCR for detection of NPC blood circulating LMP1 30 bp deletion genetic biomarker for early diagnosis and treatment response prediction of NPC patients. The analytical and diagnostic evaluation and treatment response prediction were conducted using NPC patients’ whole blood (WB) and tissue samples and non-NPC cancer patients and healthy individuals’ WB samples. The assay was able to detect as low as 20 ag DNA per reaction (equivalent to 173 copies) with high specificity against broad reference microorganisms and archive NPC biopsy tissue and FNA samples. The diagnostic sensitivity and specificity were 83.3% and 100%, respectively. The 30 bp deletion genetic biomarker was found to be a good prognostic biomarker associated with overall clinical outcome of NPC WHO type III patients. This sensitive and specific assay can help clinicians in early diagnosis and treatment response prediction of NPC patients, which will enhance treatment outcome and lead to better life-saving.

show abstract

Section: Analytical Sensitivity and Specificity Of The Developed Innoprimers-duplex Qpcr Assaymentioning

confidence: 50%

Section: Discussionmentioning

confidence: 89%

Design of InnoPrimers-Duplex Real-Time PCR for Detection and Treatment Response Prediction of EBV-Associated Nasopharyngeal Carcinoma Circulating Genetic Biomarker

et al. 2021

View full text Add to dashboard Cite

show abstract

“…According to the manufacturer's instructions, viral DNA was extracted from clinical specimens of FRI patients by using a viral DNA/RNA nucleic acid extraction and purification kit (Xi'an Tianlong Technology Co., Ltd, China) and stored at −80 °C until testing. The preliminary screening of the eight FRI-associated viruses mentioned above was performed by real-time PCR 53 . The 994 nt NS1-VP1u region (positions 2117 to 3110 in NC_000883.2) and the near complete coding B19V genome (positions 616 to 5174 in NC_000883.2) were amplified by nested PCR from the B19V-positive specimens by using Platinum PCR SuperMix (Invitrogen, USA).…”

Section: Methodsmentioning

confidence: 99%

The epidemiological and genetic characteristics of human parvovirus B19 in patients with febrile rash illnesses in China

Jiang,

Qiu,

Zhou

et al. 2023

Sci Rep

Self Cite

View full text Add to dashboard Cite

To understand the epidemiological and genetic characteristics of B19V, a multiple-province surveillance of patients with febrile rash illnesses (FRIs) were conducted in China during 2009 ~ 2021. The clinical specimens of 3,820 FRI patients were collected and tested for B19V DNA. A total of 99 (2.59%) patients were positive for B19V, and 49 (49.49%) were children under 5 years old. B19V infections occurred throughout the year without obvious seasonal pattern. Ten NS1-VP1u sequences and seven genome sequences were obtained in this study, identified as subgenotype 1a. Combined with the globally representative genome sequences, no temporal and geographic clustering trends of B19V were observed, and there was no significant correlation between B19V sequences and clinical manifestations. The evolutionary rate of the B19V genome was 2.30 × 10–4 substitutions/site/year. The number of negative selection sites was higher than that of positive selection sites. It was the first to comprehensively describe the prevalence patterns and evolutionary characteristics of B19V in FRI patients in China. B19V played the role in FRI patients. Children under 5 years old were the main population of B19V infection. Subgenotype 1a was prevalent in FRI patients in China. B19V showed a high mutation rate, while negative selection acted on the genome.

show abstract

“…Более того, Cui A. и соавт. описали методику ПЦР, по зволяющую одновременно выявлять вирус кори, вирус краснухи, энтеровирус человека, вирус ветряной оспы, вирус денге, парвовирус человека В19, вирус Эпштейна Барр и вирус герпеса человека 6 типа [50].…”

Section: патогенезunclassified

Current approaches and prospects for the development of laboratory diagnosis of measles

Nosova

Bogoslovskaya

Shipulin

2023

CMAC

View full text Add to dashboard Cite

Measles virus causes an acute infectious disease with high contagiousness. It is possible to limit the spread of measles virus only with a sufficiently wide coverage of the population by vaccination. Despite the success of measles elimination programs, many countries have seen an increase in the incidence of measles in recent years, making early diagnosis increasingly important. The importance of laboratory diagnosis is related to the difficulties of clinical differential diagnosis of measles in the early stages of the disease. This review is devoted to an analysis of existing methods for diagnosing measles. It demonstrates the limitations of the most commonly used method, the enzyme immunoassay, and the need to develop and implement alternative diagnostic methods. Particular attention in the review is paid to molecular diagnostic methods, the sensitivity of which is reviewed for different types of biological sampled at different stages of the disease. Characteristics of the measles virus that are of key importance in the development of PCR tests are described. Studies evaluating the significance of introducing PCR in the routine diagnosis of measles are presented. The main advantages of molecular methods are the possibility of early detection of the virus and the possibility of simultaneous detection of several pathogens, which allows differential diagnosis of diseases with a similar clinical presentation. The development and implementation of rapid and accurate approaches based on molecular diagnostic methods into the health care system is an urgent need in the implementation of global and local programs for the elimination of measles.

show abstract

Development of a multiplex one-step real-time RT-PCR assay for the simultaneous detection of eight viruses associated with febrile rash illnesses

Cited by 5 publications

References 25 publications

Design of InnoPrimers-Duplex Real-Time PCR for Detection and Treatment Response Prediction of EBV-Associated Nasopharyngeal Carcinoma Circulating Genetic Biomarker

Design of InnoPrimers-Duplex Real-Time PCR for Detection and Treatment Response Prediction of EBV-Associated Nasopharyngeal Carcinoma Circulating Genetic Biomarker

The epidemiological and genetic characteristics of human parvovirus B19 in patients with febrile rash illnesses in China

Current approaches and prospects for the development of laboratory diagnosis of measles

Contact Info

Product

Resources

About