Different profiles of chemokines, cytokines and cell growth factors in plasma samples from patients with leprosy, leprosy reactions and households contacts

BACKGROUND Leprosy is a highly neglected disease that is considered a serious public health problem in many countries. This illness is characterised by a variety of clinical and histopathological manifestations that are related to the patient immune response. OBJECTIVES This work aimed evaluate the profile of circulating immune mediators in the plasma from patients classified clinically as paucibacillary (PB), multibacillary (MB), households contacts (HHC), type1 leprosy reaction (T1R), type2 leprosy reaction (T2R) and control individuals without medical history of leprosy (CTL). METHODS To assessment of the plasma immune mediators was used multiplex microbeads immunoassay "Luminex". FINDINGS The results showed that patients (PB) had a regulatory-biased profile, while MB revealed a pro-inflammatory trend of highly expressed biomarkers. HHC display conspicuously increased levels in the plasma of the chemokines (CCL2, CCL3, CCL4, CCL5 and CXCL8), pro-inflammatory cytokines (IFN-γ,TNF and IL-1β), modulating cytokines (IL-9 and IL-1Ra) and growth factors (PDGF, G-CSF and IL-2). Interestingly, HHC displayed superior production of IFN-γ as compared to other leprosy groups, indicating a putative protective role for this cytokine during chronic Mycobacterium leprae exposure. MAIN CONCLUSION Further investigations are currently underway to elucidate the potential of these mediators as biomarkers applicable to the diagnosis/prognosis of leprosy and also T1R and T2R leprosy reactions.

Detection of aqueous humor cytokines in diabetic retinopathy and its clinical significance / 中华实验眼科杂志

Objective:To detect the concentration of various cytokines in aqueous humor of patients with diabetes retinopathy (DR) with Luminex liquid chip, and analyze the relationship between the cytokines and the occurrence and development of DR.Methods:A cross-sectional study was conducted.Sixty-three DR patients (97 eyes) treated with anti-vascular endothelial growth factor (VEGF) drugs in the First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine from March 1, 2019 to December 31, 2019 were enrolled as DR group, including 38 nonproliferative DR (NPDR) eyes in NPDR group and 59 proliferative DR (PDR) eyes in PDR group, 39 eyes in photocoagulation group and 58 eyes in non-photocoagulation group.Twenty-seven patients (31 eyes) hospitalized for cataract surgery at the same time were collected as the control group.Aqueous humor was extracted during the operation, and Luminex liquid chip was used to detect the concentrations of vascular endothelial growth factor-A (VEGF-A), placental growth factor (PLGF), platelet-derived growth factor (PDGF)-AA, PDGF-BB, angiopoietin-like protein 4 (ANGPTL4), interleukin-6 (IL-6), IL-8, IL-1β, monocyte chemotactic protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1) and tumor necrosis factor-α (TNF-α) in aqueous humor.The concentrations of various cytokines of different groups were compared, and the correlation among various aqueous humor cytokines was analyzed by Spearman rank correlation analysis.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of First Affiliated Hospital of Guangzhou University of Chinese Medicine (No.Y[2019]230). Written informed consent was obtained from each patient.Results:The concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8, MCP-1 and ICAM-1 in DR group were significantly higher and the concentration of IL-1β was significantly lower than those of control group ( Z=-4.747, -5.164, -3.373, -8.062, -4.535, -5.954, -5.098, -3.228, -5.954, all at P<0.01). The concentrations of VEGF-A, PLGF, PDGF-AA, ANGPTL4, IL-6, IL-8 and MCP-1 of the photocoagulation and non-photocoagulation groups were higher and the concentration of IL-1β was significantly lower than those in the control group (all at P<0.017). The concentration of ICAM-1 in the photocoagulation group was significantly higher than that in the control group ( P<0.017). The concentrations of PLGF, PDGF-AA and ANGPTL4 of PDR group were higher than those of NPDR group, and the differences were statistically significant ( Z=-2.291, -3.396, -2.276, all at P<0.05). VEGF-A was positively correlated with the other cytokines except ICAM-1 ( rs=0.237-0.540, all at P<0.05). ANGPTL4 was positively correlated with the other cytokines except IL-1β ( rs=0.361-0.733, all at P<0.01). Conclusions:The occurrence and development of DR are closely related to VEGF family, PDGF family, ANGPTL family and inflammatory factors.The concentrations of PLGF, PDGF-AA and ANGPTL4 are higher in PDR eyes.There are close and complex relationships among a variety of cytokines in the eyes of DR patients.

Bioactive ingredients in activated platelet-rich plasma derived from cord blood and adult blood via Luminex technology: a comparative study / 中国输血杂志

【Objective】 To compare the bioactive ingredients in activated platelet-rich plasma (PRP) from cord blood and adult blood, explore its reasonable indicators reflecting the inflammatory regulation ability, in order to guide the preparation. 【Methods】 PRP was prepared and activated from 63 healthy adults (31 males, 32 females) and 61 neonates (30 males, 31 females), and 20 cytokines were measured using Luminex technology for assessing the age- and sex-based bioactive differences of PRP. High-sensitivity C-reactive protein(hs-CRP), procalcitonin and MMPs/TIMPs from each sample were measured for their correlations with the 10 inflammation-related cytokines. 【Results】 The activated cord blood PRP released 10 growth factors and chemokines more than the adult blood PRP, whereas IGF-1, HGF and 8 pro-inflammatory cytokines lower than the latter. Most cytokines of adult PRP were more in females than in males (P<0.05), except for IGF-1 and HGF, which showed no difference by gender. Compared with hs-CRP and PCT, MMPs/TIMPs ratio was more closely related with the inflammation-related cytokines, which can reflect the inflammatory regulation of PRP. 【Conclusion】 Due to the lower immunocompetence and no age or gender disturbance, cord blood PRP has the rational MMPs/TIMPs ratio and more cytokines which promote the inflammation and wound healing.

Three platelet antibody detection methods: a comparative study / 中国输血杂志

【Objective】 To compare three kinds of platelet antibody detection methods used to identify alloantibodies in patients with platelet transfusion refractory(PTR). 【Methods】 The 83 samples from PTR patients were analyzed base on three different methods, including solid phase ELISA, Luminex, and capture. The sensitivity, reproducibility, and consistency of different kits were evaluated. 【Results】 A total of 71 (62 positive and 9 negative) out of 83 samples showed consistent results by three methods. The consistency between Luminex and solid phase ELISA was 95.2% (Kapp value=0.829, P<0.05), between solid phase ELISA and capture method was 85.5% (Kapp value=0.512, P<0.05), and between Luminex and capture method was 90.3% (kappa value=0.636, P<0.05). Among the 12 samples with inconsistent results, 3 cases presented positive results by capture method alone and negative by other methods, which had incompatible cross-matching results with 6 random blood donors; 5 cases with HLA antibodies showed negative results by capture method alone and positive by both Luminex and solid phase ELISA; the other 4 cases were positive in both capture and Luminex, but negative in solid phase ELISA. 【Conclusion】 The consistency of three methods was 85.5%, and each has its limitations. The capture method is rapid, economic and registered domestically, which can be used for preliminary screening.Luminex has the optimal diagnostic performance, which can be used for high-throughput and HPA/HLA antibody analysis. The solid phase ELISA is convenient. The combination of them can detect platelet antibodies effectively.

Cytokine profile of patients with leptospirosis in Sabah, Malaysia

Background: Leptospirosis is a zoonotic disease withsymptoms ranging from a mild, febrile illness to a severeform with multiorgan failure. Severe leptospirosis mayrequire medical interventions in the form of dialysis and/ormechanical ventilation and often leads to mortality. Anexaggerated host immune response—in particular, a“cytokine storm”—that causes endothelial and organdamage is associated with the disease severity andmortality.Methods: Microscopic agglutination test (MAT)-positive andMAT-negative human serum samples (n=30) from patientswith leptospirosis were obtained from the Public HealthLaboratory, Kota Kinabalu, Sabah, Malaysia and controlserum samples (n=10) were obtained from healthy studentvolunteers. We estimated the levels of IL-1β, IL-6, IL-8, IL-10,and TNF-α in serum samples by a Luminex assay.Results: The levels of IL-6, IL-8, and IL1-β were significantlyhigher in 13% of the patients with leptospirosis compared tothe healthy controls, while the levels of IL-10 and TNF-α werenot elevated in either group.Conclusion: Our data suggest that elevated levels of IL-6, IL-8, and IL1-β may be associated with leptospirosis diseaseseverity, which requires patient follow-up for confirmation.

Evaluation of the Luminex ARIES HSV 1&2 Assay and Comparison with the FTD Neuro 9 and In-house Real-Time PCR Assays for Detecting Herpes Simplex Viruses

BACKGROUND: Human herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2) are responsible for a plethora of human diseases, of which cutaneous and mucocutaneous infections are the most prevalent. In its most severe form, HSV infection can cause meningitis/encephalitis. We compared the Luminex ARIES HSV 1&2 assay (Luminex Corp., Austin, TX, USA), an automated sample-to-result molecular solution, to two non-automated HSV DNA assays. METHODS: A total of 116 artificial controls were used to determine the analytical performance of the ARIES assay. Controls were prepared by spiking universal transport medium (UTM) and cerebrospinal fluid (CSF) samples from patients who tested negative for HSV by an in-house HSV-1 and -2 DNA assay with reference materials (SeraCare Life Sciences, MA, USA; ZeptoMetrix Corp., MA, USA). Another 117 clinical samples were then used to compare the clinical performance of the ARIES assay with those of an in-house assay and the FTD Neuro 9 assay (Fast Track Diagnostics, Junglinster, Luxembourg). RESULTS: The analytical sensitivity (95% limit of detection) of the ARIES assay was 318 copies/mL (UTM samples) and 935 copies/mL (CSF samples) for HSV-1 strain 96 and 253 copies/mL (UTM samples) and 821 copies/mL (CSF samples) for HSV-2 strain 09. No cross-reactivity was observed in samples spiked with 14 non-HSV microorganisms. Compared with the reference result (agreement between the in-house and FTD Neuro 9 results), the ARIES assay had overall concordance rates of 98.2% (111/113) and 100% (113/113) for HSV-1 and HSV-2, respectively. CONCLUSIONS: The ARIES assay appears to be an excellent alternative for rapid detection and differentiation of HSV in skin and genital infections, meningitis, and encephalitis.

A fluorescent bead-based multiplex PCR assay for the simultaneous detection of seven viral diseases endemic in Africa / 中华实验和临床病毒学杂志

Objective@#To establish a fluorescent bead-based multiplex assay for the simultaneous detection of seven viral diseases endemic in Africa.@*Methods@#The genomic sequences of the viral pathogens causing Rift valley fever, Yellow fever, Marburg virus disease, Ebola virus disease, Lassa fever, Crimean-Congo hemorrhagic fever and Chikungunya fever were compared, PCR detection target fragments were selected, and amplification primers and hybrid probes were designed. The reference samples of related pathogens were prepared by chemical synthesis of DNA and in vitro transcription RNA. The sensitivity and stability of the detection method were evaluated. The specificity was evaluated by testing 30 samples of suspected dengue fever, and hantavirus diseases, and 32 healthy human blood samples.@*Results@#The fluorescent bead-based multiplex assay could specifically detect the corresponding pathogen, the detection limit was at a range of 102-105 copies/ μl, the specificity was 100%, and the intra-assay coefficient of variation was below 12%, and the inter-assay coefficient of variation was below 15%.@*Conclusions@#A fluorescent bead-based multiplex PCR assay for the simultaneous detection of seven viral diseases endemic in Africa was established, which may provide a new choice for the screening of suspected infectious diseases.

The miss rate analysis of anti-HLA antibodies detection for kidney transplantation preoperative recipients with sensitization history based on luminex xMAP / 中华器官移植杂志

Objective To interrogate the detection of anti-HLA antibodies using two methods of Luminex xMAP,and to compare their detection capacity and to analyze their misdetection rate for initial screening,providing more accurate results in clinical practice.Methods 214 serum samples from recipients with a history of sensitization before renal transplantation were collected and detected by LM (LABScreen Mixed) and LSA (LABScreen Single Antigen) respectively on the Luminex xMAP platform.Results For the LM detection,the positive rates of anti-HLA class Ⅰ and Ⅱ were 50.9％ and 23.4％ respectively,which were lower than those used by the LSA detection (58.9％ and 46.7％ respectively).The difference had statistical significance (P ＜ 0.05).The sensitivity,specificity,miss rate and mistake rate of anti-HLA class Ⅰ and Ⅱ[detection were 80.2％,90.9％,19.8％,9.1％ and 49.0％,99.1％,51.0％,0.9％ respectively.The missed detection gene with the highest rate was Cw * 17:01,B * 15:12,B * 45:01 for anti-HLA class Ⅰ and DPA1 * 01:03,DPB1 * 06:01,DPA1 * 01:03,DPB1 * 01:01 for anti-HLA class Ⅱ.The highest MFI value was 10603 and 3659.For the recipients with only blood transfusion history or pregnancy history,LM and LSA detection showed no statistically significant difference when detecting anti-HLA class Ⅰ antibodies,but had statistically significant difference when testing anti-HLA class Ⅱ antibodies.For the patients with history of both blood transfusion and pregnancy history,LM and LSA showed no significant difference in the detection of anti-HLA class Ⅰ antibodies and anti-HLA class Ⅱ antibodies.The miss rate of anti-HLA class Ⅰ antibody detection was lower than that of anti-HLA class Ⅱ antibody detection.Conclusion LSA detection has the higher sensivity and lower miss rate than LM detection.In the light of the disadvantage of LM detection as diagnostic preliminary screening,it is suggested that LSA detection should be used directly for the recipients with a history of sensitization.By this process optimization,it is more likely to cause missent inspection and the occurrence of rejection,as well as a poor long-term survival rate.

Cuantificación de la quimiocina CCL5 en pacientes con diabetes mellitus tipo 2 y/o periodontitis cronica: Estudio preliminar

l propósito del presente estudio fue cuantificar la presencia de la quimiocina CCL5 (RANTES) en Líquido Crevicular Gingival (LCG) de pacientes con Periodontitis Crónica (PC) y/o Diabetes Mellitus tipo 2 (DM2). Se realizó un estudio comparativo, transversal en 40 pacientes. Se tomó LCG de bolsas periodontales y surcos gingivales de 4 grupos de pacientes (10 por grupo de estudio), se excluyó a los pacientes que recibieron tratamiento periodontal, antibiótico y antiinflamatorio 6 meses anteriores al estudio o cursaron con alguna enfermedad sistémica distinta a DM2. Las concentraciones de CCL5 se determinaron mediante ensayos LUMINEX de selección magnética. Se realizó estadística descriptiva, prueba ANOVA de una vía, T de student y correlación de Pearson. La cuantificación de CCL5 fue mayor en los pacientes que presentaron ambas enfermedades, seguidos del grupo con solo PC, los sanos y el grupo con solo DM2. No se encontró diferencia significativa entre los grupos y no hubo correlación entre las cuantificaciones y los indicadores glicémicos. A pesar de que las diferencias no fueron significativas, el grupo de pacientes con ambas enfermedades presentó la mayor cuantificación de CCL5. La expresión de CCL5 en LGC debe considerarse un potencial inductor de destrucción periodontal, su determinación podría ser útil para monitoreo de la salud/enfermedad de los tejidos periodontales.

he purpose of the present study was to quantify the presence of chemokine CCL5 (RANTES) in gingival crevicular fluid (LCG) in patients with chronic periodontitis (PC) and / or type 2 diabetes mellitus (DM2). A comparative cross-sectional study was conducted in 40 patients. LCG was taken from periodontal pockets and gingival grooves from 4 patient groups (10 per study group); patients who received periodontal, antibiotic and anti-inflammatory treatment 6 months prior to the study or who had systemic disease other than DM2 were excluded. Concentrations of CCL5 were determined by LUMINEX® assays. Descriptive statistics, one-way ANOVA, Student's T, and Pearson's correlation were performed. The quantification of CCL5 was higher in the patients who presented both diseases, followed by the group with only PC, healthy and the group with only DM2. No significant difference was found between groups and there was no correlation between quantifications and glycemic indicators. Although the differences were not significant, the group of patients with both diseases had the highest CCL5 quantification. The expression of CCL5 in LGC should be considered as a potential inducer of periodontal destruction, its determination could be useful for monitoring the health/disease of periodontal tissues.

Clinical observation on the removal of donor specific antibody by double filtration plasmapheresis in renal transplant recipients / 器官移植

Objective To investigate the effect of double filtration plasmapheresis (DFPP) upon the removal of donor specific antibody (DSA) in highly sensitized recipients with renal transplantation. Methods Four highly sensitized recipients undergoing renal transplantation received 7 cycles of DFPP. Luminex technology was adopted to monitor the changes of DSA. Clinical efficacy, incidence of acute rejection and adverse reactions were observed. Results After DFPP, the DSA MFI [1036 (0-4113)] was significantly declined than that before treatment [6446 (2999-12905), Z= -2.503, P=0.012]. No hyperacute rejections occurred in four highly sensitized recipients undergoing renal transplantation.Acute rejection was noted in one case, which was mitigated by postoperative DFPP and adjustment of immunosuppressive agents. During postoperative follow-up, the function of transplant kidney was normal and no rejection reactions occurred. The level of albumin was decreased after DFPP. Conclusions DFPP can effectively remove the DSA in the recipients.It is an efficacious and safe approach to prevent the incidence of acute rejections in highly sensitized recipients after renal transplantation.

Application of Luminex liquid chip technology in diagnosis and monitoring for patients with Crohn's disease / 局解手术学杂志

Objective This study aimed to evaluate the clinical significance of detecting cytokine expression with Crohn's disease(CD) by Luminex liquid chip.Methods A total of 76 patients with CD and 50 healthy volunteers as healthy control group were recruited in this study.The expression of cytokines IL-2,IFN-γ,TNF-α,IL-4,IL-6,IL-10 and IL-17A were detected by Luminex liquid chip according to the instruction of MILLIPLEX MAP Kit.Enzyme-linked immunosorbent assay(ELISA) was used to detect the levels of anti-saccharomyces cerevisiae antibodies(ASCA).Then the relationship and differences among these groups were analyzed.Results Except IL-17A,the level of IL-2,IFN-γ,TNF-α,IL-4,IL-6,IL-10 was higher in the CD group than those in control group.The serum levels of IFN-γ,TNF-α and IL-6 in patients with active stage of CD group were significantly higher than those with remission stage and healthy control group.The level of TNF-α and IL-6 in moderate and severe stages were higher than those in slight stage.The serum level of ASCA had significant difference between CD group and control group.Compared with ASCA negative group,the ASCA positive patients have a higher level of IL-6 and TNF-α.ConclusionThe cytokines like IL-6 and TNF-α is correlated with CD severity,which is important for inflammatory activity and progression of CD.Altogether,these results demonstrated that the dynamic change of cytokines had a clear relativity in stage of CD,which is very imperative for diagnosis and clinical appraisal of CD.

Pathogenic characteristics and molecular typing of Salmonella strains isolated from diarrhea patients in Wuxi City, Jiangsu Province, China / 中国人兽共患病学报

We analyzed the pathogenic characteristics of Salmonella strains isolated from diarrhea patients in Wuxi City,Jiangsu Province,China and compared the differences among pulse field gel electrophoresis (PFGE) patterns of main serotype strains,so as to provid scientific basis for disease control.After biochemical identification of the Salmonella strains isolated from infectious diarrhea patients in Wuxi in 2015,drug susceptibility test,serotyping and PFGE were applied to analyze these strains.Results showed that a total of 32 Salmonella strains were detected from 756 diarrhea specimens with a positive rate of 4.23 ％.The infection occurred more frequently between May and October and adults aged more than 60 years old affected mostly.There was no significant difference between genders in infected population.The drug susceptibility test indicated that the antibiotic resistance rate of these Salmonella strains to ampicillin (56.25 ％) was the highest,and to ciprofloxacin(6.25 ％)and Ceftazidime (6.25％) were the lowest.The 32 Salmonella strains belonged to 11 serotypes,and S.enteritidis(31.25％)and S.typhimurium(21.88％) were the predominant serotypes.PFGE showed that the pattern similarity of all S.enteritidis was more than 85 ％;PFGE patterns of S.typhimurium were different.In conclusion,the infection of Salmonella from diarrhea patients in Wuxi City had obvious season and age specific distribution,and the most prevalent serotype of Salmonella was the S.enteritidis.It is necessary to strengthen the surveillance of Salmonella concurrently in food and environment.

Comparison of the Luminex xTAG Respiratory Viral Panel Fast v2 Assay With Anyplex II RV16 Detection Kit and AdvanSure RV Real-Time RT-PCR Assay for the Detection of Respiratory Viruses

BACKGROUND: The accurate and rapid identification of the causative viruses is important for the timely diagnosis and management of respiratory infections. Multiplex molecular diagnostic techniques have been widely adopted to detect respiratory viruses. We compared the results of a newly upgraded, multiplex, molecular bead-based respiratory viral panel (RVP) assay with the results of Anyplex II RV16 detection kit and AdvanSure RV real-time RT-PCR assay. METHODS: We tested 254 respiratory specimens and cultured viral strains using the Luminex xTAG RVP Fast v2 assay (Luminex Molecular Diagnostics, Canada) and Anyplex II RV16 detection kit and compared the results. Specimens showing discordant results between the two assays were tested with a AdvanSure RV real-time RT-PCR assay. RESULTS: Of the 254 respiratory specimens, there was total agreement in the results between the xTAG RVP Fast v2 assay and the other real-time PCR assay in 94.1–100% of the specimens. The agreement levels were relatively low (94.1–97.6%) for specimens of adenovirus, coronavirus NL63, and parainfluenza type 3. In comparison to the other assay, the xTAG RVP Fast v2 assay detected a higher number of parainfluenza type 3 (4 cases) and metapneumovirus (9 cases). CONCLUSIONS: The xTAG RVP Fast v2 assay showed comparable capabilities compared with the other assays; it will be useful for identifying respiratory viral infections in patients with respiratory symptoms. Clinicians should be aware of the characteristics of the assays they use, since different assays show different detectability for each virus.

Effect of anti-HLA antibody detected by Luminex testing on clinical prognosis of renal transplantation ;recipients / 器官移植

Objective To investigate the value of anti-human leukocyte antigen (HLA)antibody level detected by Luminex testing in predicting clinical prognosis of renal transplantation recipients. Methods A total of 1 105 patients scheduled to undergo renal transplantation (354 successfully undergoing renal transplantation)in the 181st Hospital of People's Liberation Army from June 2013 to November 2015 were selected. The serum samples were collected from 1 923 cases before and after renal transplantation. The positive rate and fluorescent intensity of anti-HLA antibody were detected by Luminex testing before and after renal transplantation. The renal function of recipients was also evaluated after renal transplantation. Results Prior to renal transplantation,51.0%(546/1 071)of serum samples were positive for anti-HLA antibody,including 26.0%(279/1 071)positive for anti-HLAⅠantibody,24.9%(267/1 071)positive for anti-HLAⅡantibody and 11.4% (122/1 071 )positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies. Among 354 patients undergoing renal transplantation,59 (17%)were positive for anti-HLA antibody after renal transplantation,including 25 (4 newly positive after surgery)positive for anti-HLAⅠantibody,15 (1 newly positive after surgery)positive for anti-HLAⅡantibody and 19 (4 newly positive after surgery)positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies. During subsequent follow-up,13 patients positive for anti-HLAⅠantibody,5 positive for anti-HLAⅡantibody and 1 1 positive for both anti-HLA Ⅰ and anti-HLA Ⅱ antibodies developed transplant kidney dysfunction. All patients newly positive for anti-HLA antibody after renal transplantation presented with transplant kidney dysfunction. Conclusions Luminex testing can perform dynamic detection of the positive rate of anti-HLA antibody,which is important in predicting clinical prognosis of recipients after renal transplantation.

Preliminary application of Tem-PCR combined with luminex for detection of four common respiratory vi-ruses / 医学研究生学报

Objective Respiratory viruses are the most common pathogens to cause respiratory tract infection in infants and children.The aim of the study was to establish a luminex-based molecular assay for rapid detection of four kinds of common respiratory viruses and provide measures for effective prevention and control . Methods 120 throat swab samples from patients with acute respiratory tract infection were collected in our hospital as disease group.30 normal specimens were used as control group .Specific up-stream and downstream primers , hybridization probes and super prim-ers were designed on the basis of conserved sequences of Influenza A and B viruses( FluA, FluB), respiratory syncytial virus types A and B ( RSVA, RSVB ) from available respiratory-virus sequence data-base.Recombinant plasmid and in vitro transcription RNA positive reference substances were established respectively .The testing sys-tem of Tem-PCR combined with luminex xMAP was built by amplification and optimization of hybridization .Comparative analysis were made between the detection results of the above method and those of single viral gene real -time PCR assay and luminex xTAG assay re-spectively. Results Rapid molecular assay was established to specifically detect the four kinds of respiratory viruses (FluA, FluB, RSVA and RSVB) with the sensitivity of 10 copies/μL.Rapid molecular assay and single viral real-time PCR assay were utilized to de-tect the throat swabs ( n=120 ) from suspected patients , the positive result of the former was 31 .7% ( 38/120 ) and the latter was 29.2%(35/120).The consistency test result indicated the two methods were consistent without a significant difference (k>0.7). Several samples were detected by luminex xTAG assay simultaneously , in which good consistency and significant difference were found in two assays by statistical analysis (k>0.6). Conclusion Preliminary clinical application has confirmed the novel molecular assay is sensitive, specific and rapid in simultaneous detection of FluA , FluB, RSVA and RSVB respiratory viruses , which provides experi-mental basis for accurate diagnosis of infected pathogens at early clinical stage .

Analysis of human papillomavirus infection in non-small cell lung cancer / 国际肿瘤学杂志

Objective To detect the infection of human papillomavirus (HPV)in patients with non-small cell lung cancer (NSCLC)and explore the relationship between HPV infection and clinicopathological features.Methods HPV detection and genotyping in 156 cases of NSCLC were performed using a new liquid chip based on Luminex.Patient clinical characteristics were also recorded,and the relationship between HPV infection and clinicopathological features was studied.Results Of the 156 tumor-DNA samples tested,40 (25.6%)cases showed presence of HPV-DNA,of which 37 cases were of a high-risk HPV type (16,18,33, 58).The differences were statistically significant between the HPV-positive and HPV-negative groups in gender (χ2 =4.387,P =0.036),smoking (χ2 =8.130,P =0.004),histologic type (χ2 =4.075,P =0.044)and lymph node metastasis (χ2 =7.082,P =0.008).The differences were not statistically significant between the HPV-positive and HPV-negative groups in age (χ2 =0.013,P =0.910),differentiated degree (χ2 =1.727, P =0.189),clinical stages (χ2 =0.179,P =0.672),distant metastasis (χ2 =3.012,P =0.083).Logistic regression analysis revealed that lymph node metastasis alone was an independent predictive factor of HPV infection in NSCLC (OR =0.384,95%CI:0.153-0.967,P =0.042),and gender (OR =1.402,95%CI:0.522-3.769,P =0.503),smoking (OR =0.506,95%CI:0.194-1.322,P =0.506),histologic type (OR =0.393,95%CI:0.133-0.161,P =0.091)were not independent predictive factors of HPV infection. Conclusion The infection of HPV presents in part of Chinese NSCLC patients,and HPV infection may be connected with occurrence and development of lung cancer.

Luminex-based Immunoassay for Organ Transplantation / 대한이식학회지

Development of luminex-based solid phase assays enables advanced measurement of HLA antibody with sensitivity, specificity, and increasing knowledge of unacceptable antigens. In this review, we described the principle of the luminex-based assay and its current applications for organ transplantation including C1q assay, calculated panel reactive antibody, and virtual cross-matching. We also discussed the technical aspects and limitations for clinical utilization. The variables related to measurement of HLA antibody specificities and their clinical relevance remain unclear, therefore the interpretation of results requires comprehensive knowledge and clinical information in critical cases.

Performance of LIFECODES HLA-DQB1 Typing Kit Using Luminex Platform in Koreans

Intermediate-resolution HLA-DQ typing has gained importance in organ transplantation recently. We evaluated the performance of the LIFECODES HLA-DQB1 typing kit (Immucor, USA) using sequence-specific oligonucleotide (SSO) probe and Luminex platform (Luminex Corp., USA) on 100 samples tested by sequence-based typing (SBT) using the AlleleSEQR HLA-DQB1 kit (Abbott Molecular, USA) in Korean individuals. No sample showed ambiguity in the assignment of 4-digit HLA-DQB1 allele with the LIFECODES HLA-DQB1 SSO typing kit, and the results were fully concordant with those of high-resolution typing of AlleleSEQR HLA-DQB1 SBT up to 4-digit level. Three samples required adjustment of false reactions (3/100, 3.0%): two samples with DQB1*03:03/*06:01 showed false-positive result in probe 253, and 1 sample with DQB1*04:02/*05:02 showed false-negative result in probe 217. We tested an additional sample with DQB1*03:03/*06:01, which showed same false-positivity in probe 253 and 2 samples with DQB1*04:02/*05:02, which showed no false reaction. The false reactions did not result in ambiguity or change in the HLA allele assignment. We could assign HLA-DQB1 alleles to 4 digit-level without ambiguity, with 100% concordance with the SBT results. Thus, LIFECODES HLA-DQB1 SSO typing kit showed good performance for intermediate-resolution HLA-DQB1 typing in clinical laboratory for organ transplantation in Koreans.

Analysis of the effects of miRNAs on the expression of cytokines by THP-1 cells using Luminex xMAP technology / 中华微生物学和免疫学杂志

Objective To investigate the effects of a miRNA family member, let-7e, and a combi-nation of miR-106b and miR-20a on the expression of cytokines by THP-1 cells with Luminex xMAP technol-ogy.Methods The efficiency of transfection was evaluated by immunofluorescence assay after transfecting THP-1 cells with micrONTM mimic negative control (Cy3) for 24 h, 36 h and 48 h.The three miRNA mim-ics (let-7e, miR-106b and miR-20a) were respectively used to transfect the THP-1 cells for 24 h, 36 h and 48 h and the expression of each miRNA was analyzed by qRT-PCR analysis for screening out the optimal transfection time.The transfected THP-1 cells were stimulated with1 mg/L of LPS for 1 h.The Luminex xMAP technology was used to detect the expression of IL-8, interferon-inducible protein-10 (IP-10), mono-cyte chemotactic protein 1 (MCP-1), IL-1α, IL-6, IL-10, TNF-α, IFN-αand IFN-βin the supernatants of cell culture.A statistical analysis was performed to analyze the data obtained by using SPSS16.0 software. Results More than 90% of the transfected THP-1 cells were labeled with red fluorescence.The optimal transfection times for let-7e mimic and miR-106b/miR-20a mimics were 48 h and 24 h, respectively.Com-pared with the corresponding negative control (NC), the expression of IL-8, IP-10 and MCP-1 by THP-1 cells were enhanced after the transfection with let-7e mimic, but were inhibited after the co-transfection with miR-106b and miR-20a mimics.Conclusion The expression of IL-8, IP-10 and MCP-1 were enhanced in let-7e transfected THP-1 cells, but were inhibited in miR-106b and miR-20a co-transfected THP-1 cells.

Neonatal Alloimmune Thrombocytopenia Caused by Anti-HLA-A2 Alloantibodies Determined by Luminex Single Antigen Bead Assay

Neonatal alloimmune thrombocytopenia (NAIT) occurs when maternal alloantibodies react to antigens expressed on fetal platelets, which is mainly platelet-specific alloantigen or HLA, resulting in their immune destruction. Here, we described a patient who suffered from NAIT caused by anti-HLA-A2 antibody. Sera from the mother and the newborn were screened for human platelet antigen-specific antibodies and HLA antibodies by ELISA, and HLA antibodies were detected in both of them. The antibody specificity was identified as anti-HLA-A2 by Luminex single antigen bead assay. HLA typing results showed that patient's father descended HLA-A2 antigen on the patient and the mother was HLA-A2 negative. It is most conceivable that anti-HLA-A2 alloantibody in the mother's sera crossed the placenta and subsequently caused NAIT in the case presented. The patient received platelet concentrates, oral steroid and intravenous globulin and platelet count increased to 120x10(9)/L on the 90th day of life. The Luminex single antigen bead assay used in this case is highly sensitive and specific assay to determine antibody specificity and it is faster and more convenient for routine use in clinical laboratory so that this assay could be useful to diagnose NAIT caused by HLA antibodies and treat such NAIT patients with HLA matched platelet transfusion.