1.Application of Sysmex XE-5000 automated hematology analyzer in cell counting of serous cavity effusion
Lanyun ZHUO ; Yulian PENG ; Weiye CHEN ; Xinzhong WU ; Limei MO ; Caiwen OU ; Xin PANG
International Journal of Laboratory Medicine 2014;(9):1179-1180
Objective To investigate the application of hematology analyzer in cell counting of serous cavity effusion .Methods Humoral mode of Sysmex XE-5000 automated hematology analyzer and manual microscopy were employed to perform cell counting in 85 samples of serous cavity effusion .Results Compared with the manual method and instrument method ,differences of mononu-clear cells and multinucleated cells of serous cavity effusion showed no statistical significance (P>0 .05) ,while those of leukocytes , erythrocytes were found statistical significance (P<0 .05) .Conclusion Sysmex XE-5000 automated hematology analyzer has advan-tages of simple ,stability and accuracy ,however ,its application in the effusion cell counting can not completely replace the manual microscopy .
2.Effect of high dose irradiation on thermoluminescence detector
Shuzhou RUAN ; Zhixin ZHAO ; Wenyi ZHANG ; Kaijun SU ; Xinxin PANG ; Fang WANG ; Yulian LIU ; Menghui HUO ; Changxin YAN ; Ling JIAO
International Journal of Biomedical Engineering 2019;42(4):342-345
Objective To investigate effect of high dose irradiation on the performances of thermolumines-cence detectors (LiF:Mg, Cu, P). Methods The high-dose irradiated thermoluminescence detector was annealed by a thermoluminescence annealing furnace until the annealing was completed, and then the annealed thermoluminescent detector was irradiated 0.5Gy by 137Cs irradiator to verify the accuracy of the thermoluminescentdetector. Results The thermoluminescence detector after high-dose irradiation could not be completely annealed under the temperature condition of 240 ℃, and it could be completely annealed at a high temperature as 400℃. After 0.5 Gy irradiation by 137Cs irradiator, the measurement results of the annealed thermoluminescence detector were significantly smaller, and the dose response and dispersion of the detector were also changed significantly. Conclusions After a more than 5 Gy irradiation, the crystal structure of the thermoluminescence detector has changed, and a high temperature peak above 240 ℃ has appeared, which leads to the failure to completely anneal at normal temperature. Therefore, the thermoluminescence detector can no longer be used for dosimetry after high-dose irradiation.
3.Rapid detection and genotyping of SARS-CoV-2 Omicron BA.4/5 variants using a RT-PCR and CRISPR-Cas12a-based assay.
Yunan MA ; Lirong ZOU ; Yuanhao LIANG ; Quanxun LIU ; Qian SUN ; Yulian PANG ; Hongqing LIN ; Xiaoling DENG ; Shixing TANG
Journal of Southern Medical University 2023;43(4):516-526
OBJECTIVE:
To establish a rapid detection and genotyping method for SARS-CoV-2 Omicron BA.4/5 variants using CRISPPR-Cas12a gene editing technology.
METHODS:
We combined reverse transcription-polymerase chain reaction (RT-PCR) and CRISPR gene editing technology and designed a specific CRISPPR RNA (crRNA) with suboptimal protospacer adjacent motifs (PAM) for rapid detection and genotyping of SARS- CoV-2 Omicron BA.4/5 variants. The performance of this RT- PCR/ CRISPPR-Cas12a assay was evaluated using 43 clinical samples of patients infected by wild-type SARS-CoV-2 and the Alpha, Beta, Delta, Omicron BA. 1 and BA. 4/5 variants and 20 SARS- CoV- 2-negative clinical samples infected with 11 respiratory pathogens. With Sanger sequencing method as the gold standard, the specificity, sensitivity, concordance (Kappa) and area under the ROC curve (AUC) of RT-PCR/CRISPPR-Cas12a assay were calculated.
RESULTS:
This assay was capable of rapid and specific detection of SARS- CoV-2 Omicron BA.4/5 variant within 30 min with the lowest detection limit of 10 copies/μL, and no cross-reaction was observed in SARS-CoV-2-negative clinical samples infected with 11 common respiratory pathogens. The two Omicron BA.4/5 specific crRNAs (crRNA-1 and crRNA-2) allowed the assay to accurately distinguish Omicron BA.4/5 from BA.1 sublineage and other major SARS-CoV-2 variants of concern. For detection of SARS-CoV-2 Omicron BA.4/5 variants, the sensitivity of the established assay using crRNA-1 and crRNA-2 was 97.83% and 100% with specificity of 100% and AUC of 0.998 and 1.000, respectively, and their concordance rate with Sanger sequencing method was 92.83% and 96.41%, respectively.
CONCLUSION
By combining RT-PCR and CRISPPR-Cas12a gene editing technology, we successfully developed a new method for rapid detection and identification of SARS-CoV-2 Omicron BA.4/5 variants with a high sensitivity, specificity and reproducibility, which allows rapid detection and genotyping of SARS- CoV-2 variants and monitoring of the emerging variants and their dissemination.
Humans
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COVID-19
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CRISPR-Cas Systems
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Genotype
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Reproducibility of Results
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Reverse Transcriptase Polymerase Chain Reaction
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SARS-CoV-2/genetics*
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RNA
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COVID-19 Testing