Recently, an article titled "Multi center evaluation of 7 microRNA detection reagents" was published in the Journal of Laboratory Medicine. (Laboratory Medicine, Vol. 38, Issue (11): 1075-1081). After joint evaluation and certification by 10 top tier hospitals nationwide, the precision, compliance, detection limit, and anti-interference ability of 7 miRNA test kit have shown excellent performance, which can meet the usage needs and expected uses of different regions, laboratories, and detection systems.

Project Background

MicroRNA (miRNA) is an endogenous non coding single stranded small RNA that stably exists in plasma and can regulate biological functions such as cell proliferation, differentiation, and apoptosis. Its abnormal expression is closely related to the occurrence and development of various diseases such as malignant tumors [1-2].

In recent years, circulating free miRNA has been used as a novel molecular marker for liver cancer gene detection, and its combination detection has high value in assisting early diagnosis of liver cancer. In 2011, seven miRNA specific mathematical models were established [3], making the combination detection of multiple miRNAs play a crucial role in assisting in the diagnosis and treatment of liver cancer. In the "Diagnosis and Treatment Guidelines for Primary Liver Cancer (2019 Edition)" [4], miRNA was first proposed as a novel molecular marker for liver cancer, especially for serum alpha fetoprotein negative populations.

Evaluation Object

Based on PCR technology, Dunwill Medical's "7 microRNA test kits" use a combination of 7 plasma miRNAs to accurately diagnose early liver cancer. The sensitivity is about 30% higher than the traditional liver cancer marker AFP, and it can also accurately diagnose serum AFP negative populations (sensitivity is 77.7%, specificity is 84.5%) [3]. It has obtained a Class III medical device registration certificate from the National Medical Products Administration and is widely used in clinical practice.

Evaluation Methods

Due to the influence of factors such as the testing system, personnel capabilities, and laboratory environment, the evaluation of reagent detection performance by a single laboratory has certain limitations. In order to comprehensively and objectively evaluate the detection performance of the reagent kit, this study adopted a multi-center horizontal joint evaluation method, select 10 top tertiary hospital testing centers nationwide (1. Shanghai Clinical Laboratory Center 2. Laboratory Department of the First Affiliated Hospital of China Medical University 3. Laboratory Department of the People's Hospital of Guangdong Province 4. Laboratory Department of the First Affiliated Hospital of Guangxi Medical University 5. Clinical Medical Laboratory Department of the Provincial Hospital of Shandong First Medical University 6. Medical Heritage Center of the People's Hospital of Sichuan Province 7. Laboratory Department of Zhongshan Hospital of Fudan University 8. Laboratory Department of Peking Union Medical College Hospital 9. Laboratory Department of the People's Hospital of Wuhan University 10. Naval Medical University The Laboratory Department of the Third Affiliated Hospital) and evaluates the performance of the same batch of reagents using the same batch of samples at the same time. The detection items are miR-21, miR-26a, miR-27a, miR-122, miR-192, miR-223, and miR-801, with miR-1228 as the internal reference. The evaluation parameters include precision, compliance rate, detection limit, and anti-interference ability.

Evaluation Results

1. Precision

The evaluation results of the precision of this study showed that the Ct values corresponding to the 8 miRNAs in the evaluated kit met the indicators declared by the manufacturer in the kit manual, indicating that the product has good repeatability and stability.

2. Compliance Rate

This study referred to the performance validation method in CNAS GL039-2019 document [5] and selected plasma samples from clinically diagnosed liver cancer patients and healthy controls for compliance evaluation. The total compliance rate of 10 testing centers was 100.00%, reflecting a high degree of consistency.

3. Detection limit

The evaluation results of the detection limit show that the detection rates of all 10 testing centers are 100%, confirming the high sensitivity of the reagent kit.

4. Anti interference

The evaluation results of anti-interference ability indicate that the reagent kit remains stable in samples with common interferences (such as hemolysis, hyperlipidemia, and jaundice), demonstrating strong anti-interference ability.

Conclusion

In summary, the 7 miRNA test kit has demonstrated excellent performance in precision, compliance, detection limit, and anti-interference ability, confirming their applicability and reliability in different regions, laboratories, and detection systems. This provides strong technical support for clinical diagnosis and treatment.

In addition, as the first multicenter evaluation in the field of molecular diagnosis, this study not only provides important reference for reagent selection in clinical laboratories, but also sets a benchmark for post market research and optimization of enterprise IVD products.

References:

[1] HAMMOND S M. An overview of microRNAs[J]. Advanced Drug Delivery Reviews，2015，87:3-14.

[2] SALIMINEJAD K，KHORSHID K H R，FARD S S， et al. An overview of microRNAs:biology，functions， therapeutics，and analysis methods[J]. J Cell Physiol， 2019，234(5):5451-5465.

[3] ZHOU J，YU L，GAO X，et al. Plasma microRNA panel to diagnose hepatitis B virus-related hepatocellular carcinoma[J]. J Clin Oncol，2011，29(36):4781-4788.

[4] The Medical Administration Bureau of the National Health Commission of the People's Republic of China Diagnosis and Treatment Guidelines for Primary Liver Cancer (2019 Edition) [J] Chinese Journal of Hepatology, 2020, 28 (2): 112-128 Methods [S] EP05-A2, CLSI, 2004

[5] China National Accreditation Service for Conformity Assessment CNAS GL039-2019 Molecular Diagnostic Verification Program Performance Verification Guidelines [S] Beijing: China's country of conformity assessment Family Accreditation Committee, 2019