Impact of RNA degradation on influenza diagnosis in the surveillance system

Bai, Hongyan; Zhao, Jiashen; Ma, Chunyan; Wei, Hejiang; Li, Xiyan; Fang, Qiongqiong; Yang, Peng; Wang, Quanyi; Wang, Dayang; Li, Xin

doi:10.1016/j.diagmicrobio.2021.115388

Cited by 7 publications

(7 citation statements)

References 18 publications

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“…Several drawbacks of using the RT‐PCR technique, including the questionable stability of RNA 39,40 and high detection timing of nucleic‐acid based methods, require to be compensated by methods which are rapid yet accurate, reliable and stable. Antibody‐antigen interactions have been used in clinical detection for a long time because of their high stability, unlike RNA 41 .…”

Section: Diagnostic Approachesmentioning

confidence: 99%

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

Aravind Kumar,

Aradhana,

Harleen

et al. 2023

Reviews in Medical Virology

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Studies related to clinical diagnosis and research of SARS‐CoV‐2 are important in the current pandemic era. Although molecular biology has emphasised the importance of qualitative analysis, quantitative analysis with nucleic acids in relation to SARS‐CoV‐2 needs to be clearly emphasised, which can provide perspective for viral dynamic studies of SARS‐CoV‐2. In this regard, the requirement and utilization of digital PCR in COVID‐19 research has substantially increased during the pandemic, necessitating the aggregation of its cardinal applications and future scopes. Hence, this meta‐review comprehensively addresses and emphasises the importance of nucleic acid quantification of SARS‐CoV‐2 RNA with digital PCR (dPCR). Various quantitative techniques of clinical significance like immunological, proteomic and nucleic acid‐based diagnosis and quantification, have been comparatively discussed. Furthermore, the core part of the article focusses on the working principle and advantages of digital PCR, along with its applications in COVID‐19 research. Several important applications like viral load quantitation, environmental surveillance and assay validation have been extensively investigated and discussed. Certain key future scopes of clinical importance, like mortality prediction, viral/variant‐symbiosis, and antiviral studies were also identified, suggesting several possible digital PCR applications in COVID‐19 research.

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Section: Diagnostic Approachesmentioning

confidence: 99%

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

Aravind Kumar,

Aradhana,

Harleen

et al. 2023

Reviews in Medical Virology

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“…Sample transportation and storage temperature was whole-process monitored by Testo 184 data logger. In CNIC, samples were preliminarily screened by real-time RT PCR using seasonal influenza detection primers and probes right after CNIC received the samples [18]. Positive samples were then selected for virus isolation.…”

Section: Sample Collection and Study Designmentioning

confidence: 99%

“…Nucleic acid was detected as previously described [18]. Nucleic acid was extracted from 200 µl samples using MagMAX™ CORE Nucleic Acid Purification Kit on MagMAX_Core_Flex automatic machine.…”

Section: Nucleic Acid Detectionmentioning

confidence: 99%

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Virus Isolation and Propagation from H3N2 Influenza Infected Human Clinical Samples Under Distinct Sample Storage Conditions

Bai¹,

Zhao²,

Ma³

et al. 2022

IJMB

Self Cite

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Isolation and propagation of influenza virus from Influenza Like Illness (ILI) clinical sample is essential for the surveillance of circulating virus, such as antigenic and genetic analyses, antiviral sensitivity surveillance, as well as annual influenza vaccine selection. Madin-Darby canine kidney (MDCK) cell is conventionally used for virus isolation in public health laboratories. Throat swap samples of Influenza like Illness (ILI) were collected from two sentinel hospitals and screened seasonal influenza by real-time reverse transcription polymerase chain reaction (RT-PCR). H3N2 positive samples were performed virus isolation in MDCK cells. Samples were stored under different conditions before inoculation, 1-2 days at 2-8°C, 4-5 day or 8-9 days at 2-8°C, and no less than two months at -80°C. The results showed that long term (>2 month) -80°C storage of clinical samples (15.12%) had significantly lower virus isolation rate compare to short term (1-2 days and 4-9 days) under 2-8°C storage (88.37% for 1-2 days and 52.33% for 4-9 days). For those samples stored at 4°C, the shorter of the storage time, the better of sample quality and virus activity could be obtained, resulting in higher isolation rate. This study provides evidence for influenza surveillance and sample quality control.

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“…In turn, these test results form the basis for decisions concerning their prevention and control. Regardless of sample status at the time of collection on the farm, RT-qPCR results reflect the quality and quantity of the target nucleic acid in the sample at the moment it is processed for testing in the laboratory [ 3 , 4 ]. However, between the time the sample is collected on the farm, packaged, shipped, and finally tested in the laboratory, it may have been exposed to handling conditions that adversely affect the RNA in the specimen and, therefore, the subsequent RT-qPCR test results.…”

Section: Introductionmentioning

confidence: 99%

Assessment of Strategies for Preserving Swine Viral RNA Targets in Diagnostic Specimens

Munguía-Ramírez,

Giménez-Lirola,

Zimmerman

2024

Microorganisms

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Successful downstream molecular analyses of viral ribonucleic acid (RNA) in diagnostic laboratories, e.g., reverse transcription-quantitative polymerase chain reaction (RT-qPCR) or next-generation sequencing, are dependent on the quality of the RNA in the specimen. In swine specimens, preserving the integrity of RNA requires proper sample handling at the time the sample is collected on the farm, during transport, and in the laboratory until RNA extraction is performed. Options for proper handling are limited to maintaining the cold chain or using commercial specimen storage matrices. Herein, we reviewed the refereed literature for evidence that commercial specimen storage matrices can play a role in preserving swine viral RNA in clinical specimens. Refereed publications were included if they compared RNA detection in matrix-treated vs. untreated samples. At present, the small number of refereed studies and the inconsistency in reported results preclude the routine use of commercial specimen storage matrices. For example, specimen storage matrices may be useful under specific circumstances, e.g., where it is mandatory to render the virus inactive. In a broader view, statistically sound side-by-side comparisons between specimens, viral RNA targets, and storage conditions are needed to establish if, when, and how commercial specimen storage matrices could be used in diagnostic medicine.

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Impact of RNA degradation on influenza diagnosis in the surveillance system

Cited by 7 publications

References 18 publications

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

SARS‐CoV‐2 in digital era: Diagnostic techniques and importance of nucleic acid quantification with digital PCRs

Virus Isolation and Propagation from H3N2 Influenza Infected Human Clinical Samples Under Distinct Sample Storage Conditions

Assessment of Strategies for Preserving Swine Viral RNA Targets in Diagnostic Specimens

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