Coincidence Rate Research Articles

Controlling the Fracture Location of Capsules Using the Bonding Surface Characteristics of Photocurable Resin

In recent years, self-healing research has been actively carried out to increase the life and stability of concrete structures. A representative method of concrete self-healing is a method using a bacterial substrate and a capsule. Bacteria-based self-healing is greatly influenced by the external environment, and sleeping structures are more suitable for capsule-based methods. In this study, the mechanical properties of the bonding surface of the photocurable resin are used to grow cracks in the capsule so that the self-healing capsule pops out and the healing liquid inside the capsule can be diffused efficiently. We proceeded with research to form pathways. The study analyzed the concordance rate of the bonding surface of the capsule and the judging position, and analyzed the factors affecting the concordance rate. As a result, by adjusting the number of bonding surfaces and the degree of air bubble removal, the rate of coincidence between the bonding surfaces and the fracture position was increased from 15.48% to 63.18% by a factor of 4.08.

Establishment and application of national reference panels for SARS-CoV-2 antigen detection kit

To develop a national reference panel for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen detection kit and establish a quality standard. The cultures of SARS-CoV-2 and other pathogens were collected to establish a national reference panel for SARS-CoV-2 antigen detection. The stability and homogeneity of the reference panel were evaluated. Based on World Health Organization (WHO) guidance and nucleic acid quantitative results, a quality standard reference panel was established. Currently, three generations of SARS-CoV-2 antigen national reference materials with batch numbers 370095–202001, 370095–202202, and 370095–202203 have been successfully established. These national reference panels comprised 8 positive samples, 20 negative samples, 1 repetitive sample, and 1 lower detection limit sample. The stability and homogeneity of the reference panel meet the requirements. The quality standards are as follows: the positive and negative coincidence rates are 8/8 and 20/20, respectively. The 10 test results of the medium and low-concentration repetitive reference materials should be positive, and the color rendering should be uniform (or the coefficient of variance should not be higher than 20.0%). The lower detection limit should be at least 5 × 105 U/mL (equivalent to copies/mL), and higher concentrations above the lower detection limit must be positive. A national reference panel for the SARS-CoV-2 antigen detection kit has been established. As the standard of SARS-CoV-2 antigen reagents, the reference panel has played a crucial role in the pre-marketing quality evaluation and post-marketing quality supervision in China.

Establishment and characterization of a novel indirect ELISA method based on ASFV antigenic epitope-associated recombinant protein

African Swine Fever (ASF) is an acute and highly lethal disease in pigs caused by African Swine Fever Virus (ASFV). Viral proteins have been commonly used as antigenic targets for the development of ASF diagnostic methods. However, the prokaryotic expression of viral proteins has deficiencies such as instability, insolubility, and high cost in eukaryotic situations. This study screened and verified ASFV-encoded p72, p54, and p30 protein antigenic epitopes. Subsequently, a novel antigenic epitope-associated recombinant protein was designed based on an ideal structural protein and expressed in Escherichia coli (E. coli). Western blot analysis indicated that the recombinant protein could specifically react with the monoclonal antibody (mAb) of p72 and polyclonal antibodies of p54 and p30, respectively. Next, an ASF indirect ELISA (iELISA) method was established based on the recombinant protein, which has no specific reaction with sera of other important pig viral diseases. Meanwhile, it shows a sensitivity to detecting dilutions of ASF-positive reference serum up to 1:6400. The clinical sample detection results showed a high coincidence rate of 98 % with a commercial competition ELISA kit. In conclusion, we established a novel specific, and sensitive ASF serologic detection method that opens new avenues for ASF serodiagnostic method development.

Preparation of double-system imprinted polymer-coated multi-walled carbon nanotubes and their application in simultaneous determination of thyroid-disrupting chemicals in dust samples

Dust ingestion is a significant route of human exposure to thyroid-disrupting chemicals (TDCs), and simultaneous determination of multi-contaminants is a great challenge for environmental monitoring. In this study, molecularly imprinted polymer-coated multi-walled carbon nanotubes using thyroxine as the template were synthesized for highly selective TDCs capture. This polymer was prepared by integrating the atom transfer radical polymerization using 2-(3-indol-yl)ethylmethacrylamide as the monomer with the self-polymerization of dopamine. Construction of double-system imprinted cavities could significantly improve their selective recognition performance for TDCs and the coincidence rate reached 88.5 %. The prepared polymers were applied as the solid phase extraction adsorbent to simultaneously determine 7 groups of 35 TDCs. The proposed method showed wide linear range (0.25–1000 ng L−1), low limits of detection (0.02–0.23 ng L−1) and acceptable recoveries (81.8 %–103.5 %). The occurrence and distribution of TDCs were then studied in indoor dust samples (n = 65) collected from four cities in China. We found that tetrabromobisphenol A was the predominant compound and perfluorinated compounds were the most abundant TDCs. In addition, the distribution ratio of TDCs varied between regions. This study provides an efficient technology for direct exposure assessment of multi-contaminants.

Construction of the core germplasm of yellowhorn (Xanthoceras sorbifolium Bunge) using physiological traits and SSR markers

Yellowhorn (Xanthoceras sorbifolium Bunge) is an important ecological, economic, and ornamental plant species native to China. However, the physiological diversity, genetic variation, and core germplasms have not been revealed, which hindered the breeding process of yellowhorn. Here, we evaluated the genetic diversity of 155 yellowhorn germplasms using five physiological traits. The content of flavonoids varied significantly in different germplasms, and it was positively correlated with soluble sugar content (P < 0.01). A total of 253, 336 candidate SSR were identified from the whole genome of yellowhorn. A moderate genetic diversity was found in yellowhorn based on the mean values of the number of effective alleles (Ne = 2.307), expected heterozygosity (He = 0.527), and observed heterozygosity (Ho = 0.317) with 30 pairs of polymorphic SSR markers. Population structure, Neighbor-Joining phylogeny, and principal coordinates analyses divided all genotypes into two major groups with few admixtures. Meanwhile, we extracted 34 (21.94 %) core yellowhorn germplasms based on the variation in physiological traits (14 germplasms) and genetic diversity (23 germplasms). After multiple different analyses including genetic diversity parameters, coincidence rate of range, variance difference percentage, and coefficient of variation rate of change, there was no obvious difference between core and original germplasm, which indicated that the core germplasm could represent the original germplasm well. Our study will lay a solid foundation for future conservation and management of yellowhorn germplasms and the core collection will be valuable in molecular breeding programs.

A nanobody-based blocking enzyme-linked immunosorbent assay for detecting antibodies against pseudorabies virus glycoprotein E

Pseudorabies (PR) is an acute infectious disease of pigs caused by the PR virus (PRV) and results in great economic losses to the pig industry worldwide. PRV glycoprotein E (gE)-based enzyme-linked immunosorbent assay (ELISA) has been used to distinguish gE-deleted vaccine-immunized pigs from wild-type virus-infected pigs to eradicate PR in some countries. Nanobody has the advantages of small size and easy genetic engineering and has been a promising diagnostic reagent. However, there were few reports about developing nanobody-based ELISA for detecting anti- PRV-gE antibodies. In the present study, the recombinant PRV-gE was expressed with a bacterial system and used to immunize the Bactrian camel. Then, two nanobodies against PRV-gE were screened from the immunized camel by phage display technique. Subsequently, two nanobody-HRP fusion proteins were expressed with HEK293T cells. The PRV-gE-Nb36-HRP fusion protein was selected as the probe for developing the blocking ELISA (bELISA) to detect anti-PRV-gE antibodies. Through optimizing the conditions of bELISA, the amount of coated antigen was 200 ng per well, and dilutions of the fusion protein and tested pig sera were separately 1:320 and 1:5. The cut-off value of bELISA was 24.20%, and the sensitivity and specificity were 96.43 and 92.63%, respectively. By detecting 233 clinical pig sera with the developed bELISA and a commercial kit, the results showed that the coincidence rate of two assays was 93.99%. Additionallly, epitope mapping showed that PRV-gE-Nb36 recognized a conserved conformational epitope in different reference PRV strains. Simple, great stability and low-cost nanobody-based bELISA for detecting anti-PRV-gE antibodies were developed. The bELISA could be used for monitoring and eradicating PR.

A-368 Aspergillus Galactomannan Lateral Flow Assay with Digital Reader for the Diagnosis of Invasive Aspergillosis

Abstract Background The Novel Aspergillus Galactomannan Lateral Flow Assay(Quic GM) from Dynamiker Biotechnology (Tianjin) Co., Ltd is a fluorescent immunochromatographic cassette test using a monoclonal antibody against GM with Digital Reader (Figure 1), which was rapid and easy to operate compared with the traditional GM ELISA assay. Here we conducte a comparative study between the Dynamiker Quic GM and the Platelia GM (Bio-Rad, USA) to validate the performance of the novel assay for the diagnosis of Invasive Aspergillosis (IA). Methods A total of 573 serum samples and 97 BALF samples with proven IA (Serum n = 9, BALF n = 2), probable IA (Serum n = 174, BALF n = 25), possible IA (Serum n = 98, BALF n = 16) and patients not fulfilling 2019 EORTC/MSG guideline IA criteria (control, Serum n = 292, BALF n = 54) were included. All the samples were tested in parallel using the Quic GM and Platelia GM. Results The total coincidence rate of Quic GM compared with Platelia GM was 97.91% (Kappa value was 0.95 &amp;gt; 0.75), 97.94% (Kappa value was 0.95 &amp;gt; 0.75) and 97.91% (Kappa value was 0.95 &amp;gt; 0.75) in serum samples, BALF samples and serum+BALF samples, respectively (Table 1). The concentrations generated by Quic GM and Platelia GM were linearly correlated (Serum: slope = 0.97, r2 = 0.96, P &amp;lt; 0.0001; BALF: slope = 0.95, r2 = 0.94, P &amp;lt; 0.0001) (Figure 2). Conclusion In conclusion, the Dynamiker Quic GM yielded significant consistency with Bio-Rad Platelia GM ELISA, and showed good performances for IA diagnosis, with the benefit of a relatively short handling time (around 20 min).

Erection Hardness Score in Masturbation Can Serve as a Preliminary Screening Tool for Organic Erectile Dysfunction

To explore the efficacy, in terms of sensitivity and specificity, of Erection Hardness Score (EHS) applied to masturbation in diagnosing predominantly organic erectile dysfunction (ED), compared to the actual standard, which is nocturnal penile tumescence and rigidity (NPTR). A total of 189 consecutive patients who came to our outpatient clinic from October 2019 to October 2020, reporting ED and an abridged International Index of Erectile Function (IIEF-6) score <26 were enrolled. Patients filled out two psychometric questionnaires: the IIEF-6 applied to partnered intercourse and the EHS applied to masturbation, then they underwent the NPTR test with Rigiscan for 2 consecutive nights in good sleep condition. Sensitivity, specificity, positive predictive value, and negative predictive value of EHS and IIEF-6 scales with NPTR as the standard, as well as the correlation between EHS, IIEF-6, and NPTR results were calculated. Results show that the sensitivity of the EHS scale for the diagnosis of ED was 60.0%, the specificity was 95.7%, the positive predictive value was 83.3%, the negative predictive value was 86.9%, and the coincidence rate was 86.2%. The correlation coefficient was 0.572 and the area under the ROC curve was 0.78, which means the strength of erection in masturbation as evaluated by the EHS can effectively help in discriminating between predominantly organic and apparently nonorganic ED. An EHS score of 3-4, meaning good-optimal erectile function during masturbation, decreases the likelihood of predominantly organic ED and reduces the need for further NPTR testing.

Diagnostic performance of endorectal ultrasound combined with shear wave elastography for rectal tumors staging.

This study aims to analyze the performance of endorectal ultrasound (ERUS) combined with shear wave elastography (SWE) for rectal tumor staging. Forty patients with rectal tumors who had surgery were enrolled. They underwent ERUS and SWE examinations before surgery. Pathological results were used as the gold standard for tumor staging. The stiffness values of the rectal tumor, peritumoral fat, distal normal intestinal wall, and distal perirectal fat were analyzed. The diagnostic accuracy of ERUS stage, tumor SWE stage, ERUS combined with tumor SWE stage, and ERUS combined with peritumoral fat SWE stage were compared and evaluated by receiver operating characteristic (ROC) curve to select the best staging index. From T1 to T3 stage, the maximum elasticity (Emax) of the rectal tumor increased gradually (p < 0.05). The cut-off values of adenoma/T1 and T2, T2 and T3 tumors were 36.75 and 85.15kPa, respectively. The diagnostic coincidence rate of tumor SWE stage was higher than that of ERUS stage. Overall diagnostic accuracy of ERUS combined with peritumoral fat SWE Emax restaging was significantly higher than that of ERUS. ERUS combined with peritumoral fat SWE Emax for tumor restaging can effectively distinguish between stage T2 and T3 rectal tumors, which provides an effective imaging basis for clinical decisions.

Temporal Association Among Influenza-Like Illness, Cardiovascular Events, and Vaccine Dose in Patients With High-Risk Cardiovascular Disease

Influenza-like illness (ILI) activity has been associated with increased risk of cardiopulmonary (CP) events during the influenza season. High-dose trivalent influenza vaccine was not superior to standard-dose quadrivalent vaccine for reducing these events in patients with high-risk cardiovascular (CV) disease in the Influenza Vaccine to Effectively Stop Cardio Thoracic Events and Decompensated Heart Failure (INVESTED) trial. To evaluate whether high-dose trivalent influenza vaccination is associated with benefit over standard-dose quadrivalent vaccination in reducing CP events during periods of high, local influenza activity. This study was a prespecified secondary analysis of INVESTED, a multicenter, double-blind, active comparator randomized clinical trial conducted over 3 consecutive influenza seasons from September 2016 to July 2019. Follow-up was completed in July 2019, and data were analyzed from September 21, 2016, to July 31, 2019. Weekly Centers for Disease Control and Prevention (CDC)-reported, state-level ILI activity was ascertained to assess the weekly odds of the primary outcome. The study population included 3094 patients with high-risk CV disease from participating centers in the US. Participants were randomized to high-dose trivalent or standard-dose quadrivalent influenza vaccine and revaccinated for up to 3 seasons. The primary outcome was the time to composite of all-cause death or CP hospitalization within each season. Additional measures included weekly CDC-reported ILI activity data by state. Among 3094 participants (mean [SD] age, 65 [12] years; 2309 male [75%]), we analyzed 129 285 person-weeks of enrollment, including 1396 composite primary outcome events (1278 CP hospitalization, 118 deaths). A 1% ILI increase in the prior week was associated with an increased risk in the primary outcome (odds ratio [OR], 1.14; 95% CI, 1.07-1.21; P < .001), CP hospitalization (OR, 1.13; 95% CI, 1.06-1.21; P < .001), and CV hospitalization (OR, 1.12; 95% CI, 1.04-1.19; P = .001), after adjusting for state, demographic characteristics, enrollment strata, and CV risk factors. Increased ILI activity was not associated with all-cause death (OR, 1.00; 95% CI, 0.88-1.13; P > .99). High-dose compared with standard-dose vaccine did not significantly reduce the primary outcome, even when the analysis was restricted to weeks of high ILI activity (OR, 0.88; 95% CI, 0.65-1.20; P = .43). Traditionally warmer months in the US were associated with lower CV risk independent of local ILI activity. In this secondary analysis of a randomized clinical trial, ILI activity was temporally associated with increased CP events in patients with high-risk CV disease, and a higher influenza vaccine dose did not significantly reduce temporal CV risk. Other seasonal factors may play a role in the coincident high rates of ILI and CV events. ClinicalTrials.gov Identifier: NCT02787044.

Additional role of ECC in the detection and treatment of cervical HSIL.

To probe into the additional role of ECC in the detection of cervical HSIL. The primary objective was to risk-stratify HSIL patients according to ECC so as to provide clinical suggestions for subsequent treatment. Retrospective analysis of medical records for patients with HSIL. All patients underwent both ECC and cervical biopsy. According to the results of colposcopic targeted biopsy and ECC, the patients were divided into three groups: (1) ECC negative group (those whose colposcopic targeted biopsy indicated HSIL, but ECC indicated LSIL or chronic inflammation); (2) Only the ECC positive group (those whose ECC suggested HSIL, but colposcopic targeted biopsy showed LSIL or chronic inflammation); (3) ECC and biopsy positive group (those whose ECC and targeted biopsy were both HSIL). Chi-square test was used to analyze the differences of lesion residue and biopsy results after LEEP amongst the three groups. A total of 1,146 medical records were analyzed. The diagnostic accuracy of ECC combined with colposcopic targeted biopsy for HSIL was higher than that of colposcopic biopsy alone (72.43% vs. 67.54%). When ECC indicated HSIL, the coincidence rate of ECC combined with colposcopic targeted biopsy and the histological pathology of LEEP was 86.25%, and the proportion of residual lesions after LEEP was 41.43%. When ECC and targeted biopsy both indicated HSIL, HSIL or worse lesions were confirmed in 90.68% of patients after surgery. Of these, 10.77% were confirmed as cervical invasive carcinoma. Moreover, the positive rate of LEEP resection margin and postoperative ECC in these patients was 43.48%. ECC can improve the detection rate of cervical HSIL and reduce missed diagnosis. Also ECC can help clinicians predict the proportion of residual lesions after LEEP. This provides the gynecologists with a reference for the need to increase the depth of the procedure and the need to perform ECC for the residual cervical canal.

Development of a 1-step TaqMan real-time PCR method for detection of the Bovine Group A Rotavirus

BackgroundThe purpose of this study was to develop a 1-step real-time quantitative fluorescence polymerase chain reaction (QF-PCR) method for detecting Bovine Group A Rotavirus (BRVA). The primers and probe were designed targeting the VP6 gene of BRVA. The standard substance was obtained through in vitro transcription. The primers, probe concentration, and annealing temperatures were optimized to determine the optimal system and conditions for the reaction. The specificity, sensitivity, and repeatability of the method were assessed and compared with a reported real-time QF-PCR method for clinical samples. ResultsThe results indicated that the detection method can achieve a sensitivity of 3.47 copies/μL and exhibit good specificity by exclusively detecting BRVA without cross-reactivity to other common pathogens in cattle and sheep. The standard curve exhibited a robust linear correlation, and the amplification efficiency was calculated to be 105%. The intra-group and inter-group coefficients of variation were less than 2%. A total of 96 clinical samples were tested and compared with the real-time QF-PCR method that was reported. The coincidence rate was 90.63% (87/96). Furthermore, the clinical samples revealed that the prevalence of BRV in cattle from Fujian Province was 85.42% (82/96). ConclusionThis study has successfully developed a 1-step real-time QF-PCR method for BRVA, which offers an efficient and sensitive technical support for the rapid diagnosis and epidemiological investigation of BRVA.

Identification of specific antigens between Toxoplasma gondii and Neospora caninum and application of potential diagnostic antigen TgGRA54.

Toxoplasma gondii is a zoonotic parasite that is very common in livestock. Meat products from livestock infected with T. gondii are one of the important transmission routes of toxoplasmosis. Rapid and reliable diagnosis is a prerequisite for the prevention and control of toxoplasmosis. Neospora caninum and T. gondii are similar in morphology and life history, and there are a large number of cross antigens between them, making clinical diagnosis of toxoplasmosis more difficult. In this study, immunoprecipitation-mass spectrometry (IP-MS) was used to screen for T. gondii-specific antigens, and the specific antigen was cloned and expressed in Escherichia coli. The specific antigen was then used to establish an indirect ELISA diagnostic method. A total of 241 specific antigens of T. gondii and 662 cross antigens between T. gondii and N. caninum were screened by IP-MS. Through bioinformatics analysis and homology comparison, seven proteins were selected for gene cloning and prokaryotic expression, and the most suitable antigen, TgGRA54, was selected to establish an indirect ELISA for T. gondii. Compared with the indirect immunofluorescent antibody test (IFAT), the positive coincidence rate of the ELISA based on rTgGRA54 was 100% (72/72) and the negative coincidence rate was 80.95% (17/21). The indirect ELISA method based on TgGRA54 recombinant protein was established to detect T. gondii antibodies in bovine sera, and the recombinant protein reacted well with T. gondii positive sera from sheep, mouse, and swine, indicating that the recombinant protein is a good diagnostic antigen for T. gondii.

The instantly blocking-based fluorescent immunochromatographic assay for the detection of SARS-CoV-2 neutralizing antibody.

At present, there is an urgent need for the rapid and accurate detection of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) neutralizing antibodies (NAbs) to evaluate the ability of the human body to resist coronavirus disease 2019 (COVID-19) after infection or vaccination. The current gold standard for neutralizing antibody detection is the conventional virus neutralization test (cVNT), which requires live pathogens and biosafety level-3 (BSL-3) laboratories, making it difficult for this method to meet the requirements of large-scale routine detection. Therefore, this study established a time-resolved fluorescence-blocking lateral flow immunochromatographic assay (TRF-BLFIA) that enables accurate, rapid quantification of NAbs in subjects. This assay utilizes the characteristic that SARS-CoV-2 neutralizing antibody can specifically block the binding of the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein and angiotensin-converting enzyme 2 (ACE2) to rapidly detect the content of neutralizing antibody in COVID-19-infected patients and vaccine recipients. When 356 samples of vaccine recipients were measured, the coincidence rate between this method and cVNT was 88.76%, which was higher than the coincidence rate of 76.97% between cVNT and a conventional chemiluminescence immunoassay detecting overall binding anti-Spike-IgG. More importantly, this assay does not need to be carried out in BSL-2 or 3 laboratories. Therefore, this product can detect NAbs in COVID-19 patients and provide a reference for the prognosis and outcome of patients. Simultaneously, it can also be applied to large-scale detection to better meet the needs of neutralizing antibody detection after vaccination, making it an effective tool to evaluate the immunoprotective effect of COVID-19 vaccines.

Reliability and Validity of the Chinese Version of Reflux Symptom Score

To evaluate the reliability, validity and clinical value of the Chinese version of reflux symptom score (RSS) (RSS in Chinese). This was a prospective study that contained 42 healthy volunteers and 135 possible laryngopharyngeal reflux disease (LPRD) patients. Reflux symptom index, reflux finding score, oropharyngeal pH monitoring (Dx-pH monitoring), and RSS of each patient were performed. RSS was performed again after 1 week. Confirmed LPRD patients were treated with proton pump inhibitor (PPI) for 8 weeks. And RSS was performed again after treatment. The reliability and validity of RSS was evaluated. The Cronbach's α coefficient of the Chinese version of RSS was 0.772, which indicated good internal reliability. The results of test-retest found all P values were less than 0.05, which supported good external reliability. The comparison of the results of RSS with oropharyngeal pH monitoring discovered a diagnostic coincidence rate of 83.70% and a positive predictive value of 84.96%, which showed good criterion validity. After 8 weeks treatment of PPI, RSS decreased significantly (pretreatment 84.79±42.50，post-treatment 20.11±22.82,P<0.001), indicating good responsiveness to change of RSS. The score of quality of life impact of suspected LPRD patients was obviously higher than that of healthy volunteers (t=7.153,P<0.001). All patients and volunteers agreed that RSS in Chinese can evaluate their symptoms well. RSS in Chinese had good internal and external reliability, good criterion validity and good responsiveness to change. The content and method of evaluation of RSS in Chinese was better and more comprehensive. RSS in Chinese could be a new instrument to evaluate LPRD in China.

A Recombinase-Aided Amplification Assay for the Detection of Chlamydia felis.

Chlamydia felis is an important zoonotic agent for humans and various animals. A recombinase-aided amplification (RAA) assay was developed for detecting C. felis. RAA can be performed in a closed tube at 39°C within 30 min. The detection limit was 10.6 copies of the C. felis plasmid DNA per reaction. No positive signals for other pathogens were detected. The coincidence rate of RAA and conventional PCR was 95.24% (20/21) and 100% (96/96) for positive and negative samples, respectively. The established RAA assay is a simple, rapid, highly sensitive, and specific method for detecting C. felis.

Correlation of Sysmex-XN9000 and CellaVision Advanced RBC software on anisocytosis

ABSTRACTObjectiveThis study aims to evaluate the consistency of heterogeneity degree of erythrocyte volume parameters between the blood automated analyzer Sysmex-XN9000 and the advanced red blood cell software CellaVisionDI-60.Method500 blood samples of volunteers were analyzed by Sysmex-XN9000 and CellaVision-DI60. The sensitivity, specificity, positive predictive value, negative predictive value, false positive rate, and false negative rate were evaluated. The consistency of all parameters was tested.ResultTaking the standard RBC group as the control group, the RBC parameters of the macrocytic and the microcytic group were compared. There was a statistical difference between the groups. ROC curve analysis showed that the best cutoff value of microcytic and of macrocytic affecting MCV were 4.1% and 5.7%, respectively. The best cutoff value of anisocytosis was 15.0%. The correlation coefficient between anisocytosis and red blood cell distribution width (RDW-CV) was 0.756. The sensitivity, specificity, positive predictive value and coincidence rate of anisocytosis were high. The false negative rate was 10.0%, and the false positive rate was 7.4%.ConclusionAll parameters of the degree of heterogeneity have good accuracy and consistency in the two instruments. Anisocytosis is with higher coincidence rate and positive predictive value. MIC and MAC have a good prediction on the increase or decrease of MCV. Although advanced RBC software's false negative and false positive rates are high, the red blood cell image system is more intuitive and time-saving in observing cells. Consequently, CellaVision-DI60 is suggested to combine with XN-9000 for judging the anisocytosis in daily work comprehensively.

Quantitative assessment of thenar to evaluate hand function after stroke by Bayes discriminant

BackgroundThe incidence rate of stroke or cerebrovascular accidents ranks first in China. More than 85% of stroke patients have residual upper limb motor dysfunction, especially hand dysfunction. Normalizing the rehabilitation evaluation process and standard quantitative evaluation method is a complex and key point in rehabilitation therapy. The study aimed to establish a function model based on the Bayes discriminant by measuring the thenar stiffness with shear wave elastography (SWE) to quantitatively evaluate the hand motor function of hemiplegic patients after stroke.MethodsThis study collected 60 patients diagnosed with hemiplegia after stroke from October 2021 to October 2022. Therapists used the Brunnstrom assessment (BA)scale to divide the patients into the stage. All the patients underwent the measurement of SWE examination of abductor pollicis brevis (APB), opponens pollicis (OP), flexor pollicis long tendon (FPLT), and flexor pollicis brevis (FPB) by two sonographers. The SWE change rate of four parts of the thenar area was calculated prospectively with the non-hemiplegic side as the reference, the function equation was established by the Bayes discriminant method, and the evaluation model was fitted according to the acquired training set data. Lastly, the model was verified by self-validation, cross-validation, and external data validation methods. The classification performance was evaluated regarding the area under the ROC curve (AUC), sensitivity, and specificity.ResultsThe median SWE values of the hemiplegic side of patients were lower than those of the non-hemiplegic side. According to the BA stage and SWER of APB, OP, FPLT, and FPB, our study established the Bayes discriminative model and validated it via self-validation and cross-validation methods. Then, the discriminant equation was used to validate 18 patients prospectively, the diagnostic coincidence rate was about 78.8%, and the misjudgment rate was approximately 21.2%. The AUC of the discriminant model for diagnosing BA stage I-VI was 0.928(95% CI: 0.839-1.0),0.858(95% CI: 0.748–0.969),1.0(95% CI: 1.0–1.0), 0.777(95% CI: 0.599–0.954),0.785(95% CI: 0.593–0.977) and 0.985(95% CI: 0.959-1.0), respectively.ConclusionThis Bayes discriminant model built by measuring thenar stiffness was of diagnostic value and can provide an objective basis for evaluating clinical rehabilitation.

Triplex Crystal Digital PCR for the Detection and Differentiation of the Wild-Type Strain and the MGF505-2R and I177L Gene-Deleted Strain of African Swine Fever Virus.

African swine fever (ASF) is a severe and highly contagious viral disease that affects domestic pigs and wild boars, characterized by a high fever and internal bleeding. The disease is caused by African swine fever virus (ASFV), which is prevalent worldwide and has led to significant economic losses in the global pig industry. In this study, three pairs of specific primers and TaqMan probes were designed for the ASFV B646L, MGF505-2R and I177L genes. After optimizing the reaction conditions of the annealing temperature, primer concentration and probe concentration, triplex crystal digital PCR (cdPCR) and triplex real-time quantitative PCR (qPCR) were developed for the detection and differentiation of the wild-type ASFV strain and the MGF505-2R and/or I177L gene-deleted ASFV strains. The results indicate that both triplex cdPCR and triplex qPCR were highly specific, sensitive and repeatable. The assays could detect only the B646L, MGF505-2R and I177L genes, without cross-reaction with other swine viruses (i.e., PRRSV, CSFV, PCV2, PCV3, PEDV, PDCoV and PRV). The limit of detection (LOD) of triplex cdPCR was 12 copies/reaction, and the LOD of triplex qPCR was 500 copies/reaction. The intra-assay and inter-assay coefficients of variation (CVs) for repeatability and reproducibility were less than 2.7% for triplex cdPCR and less than 1.8% for triplex qPCR. A total of 1510 clinical tissue samples were tested with both methods, and the positivity rates of ASFV were 14.17% (214/1510) with triplex cdPCR and 12.98% (196/1510) with triplex qPCR, with a coincidence rate of 98.81% between the two methods. The positivity rate for the MGF505-2R gene-deleted ASFV strains was 0.33% (5/1510), and no I177L gene-deleted ASFV strain was found. The results indicate that triplex cdPCR and triplex qPCR developed in this study can provide rapid, sensitive and accurate methods for the detection and differentiation of the ASFV B646L, MGF505-2R and I177L genes.

Development of a Quadruplex RT-qPCR for the Detection of Porcine Rotaviruses and the Phylogenetic Analysis of Porcine RVH in China.

Rotavirus A species (RVA), RVB, RVC, and RVH are four species of rotaviruses (RVs) that are prevalent in pig herds, and co-infections occur frequently. In this study, a quadruplex real-time quantitative RT-PCR (RT-qPCR) for the simultaneous detection of four porcine RVs was developed by designing specific primers and probes based on the VP6 gene of RVA, RVB, RVC, and RVH, respectively. The method showed high specificity and could only detect RVA, RVB, RVC, and RVH, without cross-reaction with other porcine viruses; showed excellent sensitivity, with a limit of detection (LOD) of 1.5 copies/µL for each virus; showed good repeatability, with intra-assay coefficients of variation (CVs) of 0.15-1.14% and inter-assay CVs of 0.07-0.96%. A total of 1447 clinical fecal samples from Guangxi province in China were tested using the developed quadruplex RT-qPCR. The results showed that RVA (42.71%, 618/1447), RVB (26.95%, 390/1447), RVC (42.92%, 621/1447), and RVH (13.68%, 198/1447) were simultaneously circulating in the pig herds, and the co-infection rate of different species of rotaviruses was found to be up to 44.01% (579/1447). The clinical samples were also detected using one previously reported method, and the coincidence rate of the detection results using two methods was more than 99.65%. The phylogenetic tree based on the VP6 gene sequences of RVH revealed that the porcine RVH strains from Guangxi province belonged to the genotype I5, which was closely related to Japanese and Vietnamese strains. In summary, an efficient, sensitive, and accurate method for the detection and differentiation of RVA, RVB, RVC, and RVH was developed and applied to investigate the prevalence of porcine RVs in Guangxi province, China. This study is the first to report the prevalence of porcine RVH in China.