The fungal bioluminescence pathway (FBP) is a metabolic pathway responsible for the generation of bioluminescence derived from fungi. This pathway utilizes caffeic acid as the substrate, generating a high-energy intermediate, and the decomposition of which yields green fluorescence with a wavelength of approximately 520 nm. The FBP is evolutionally conserved in luminescent fungal groups. Unlike other bioluminescent systems, the FBP is particularly suitable for engineering applications in eukaryotic organisms, especially in plants. Currently, metabolically engineered luminescent plants are able to emit visible light to illuminate its surroundings, which can be visualized clearly in the dark. The fungal bioluminescent system could be explored in various applications in molecular biology, biosensors and glowing ornamental plants, and even green lighting along city streets.
Luminescence ; Light ; Fluorescence ; Eukaryota ; Green Light

Objective To compare the sensitivity and accuracy of amplified luminescent proximity homogeneous assay linked immunosorbent assay (AlphaLISA) and magnetic particles-based chemiluminescence immunoassay (MP-CLIA) for detection of staphylococcal enterotoxin C (SEC) in the simulated milk samples. Methods The AlphaLISA was constructed using goat anti-SEC polyclonal antibody-coupled receptor microspheres, biotin-labeled SEC monoclonal antibody and streptavidin-coupled donor microspheres. The MP-CLIA was constructed using goat anti-SEC polyclonal antibody conjugated alkaline phosphatase, biotin-labeled anti-SEC monoclonal antibody and streptavidin conjugated magnetic beads. Results The sensitivity of AlphaLISA to detect SEC content in simulated milk samples was 4.04 ng/L, and the coefficient of variation (CV) was 1.98%~9.82%. The sensitivity of MP-CLIA was 108.19 ng/L and CV was 4.63%~20.40%. Conclusion Compared with MP-CLIA, AlphaLISA is more sensitive and accurate to detecting SEC.
Animals ; Streptavidin ; Biotin ; Luminescence ; Milk ; Antibodies, Monoclonal ; Goats ; Immunoassay/methods*

The toxin-producing bacterium Vibrio cholerae can cause severe diarrhea and has caused seven global pandemics. Traditional viable cell counts and phage plaques are commonly used to evaluate the efficacy of virulent phage clearance of V. cholerae, but these operations are time-consuming and labor-intensive, and difficult to provide real-time changes. It is desirable to develop a simple and real-time method to monitor V. cholerae during phage lysis. In this study, a luminescence-generating plasmid pBBR-pmdh-luxCDABE was transformed into three O1 serogroup drug-resistant strains of V. cholerae. The results showed that the luminescence value as a monitoring index correlates well with the traditional viable cell count method. Monitoring the number of live cells of V. cholerae by measuring the luminescence allowed real-time analysis of the number of bacteria remaining during phage lysis. This method enables repeated, interference-free, continuous multiple-time-point detection of the same sample without the time delay of re-culture or plaque formation, facilitating real-time monitoring and analysis of the interaction between the phage and the host bacteria.
Bacteriophages/genetics* ; Luminescence ; Plasmids ; Vibrio cholerae

To develop a magnetic nanoparticle chemiluminescence immunoassay (CLIA) for the determination of type Ⅰ procollagen N-terminal peptide (PINP) in human serum, we expressed a recombinant PINP-α1 protein in Corynebacterium glutamicum and used it as an immunogen to immunize BALB/c mice. We obtained three hybridoma cell lines that stably secret antibody against PINP-α1 protein. After further pairing and screening, we chose a monoclonal antibody 8C12 coupled with biotin as the capture antibody, and a monoclonal antibody 1F11 labeled horseradish peroxidase as the detection antibody. The antibodies combined with the serum samples, forming a sandwich complex which was used to detect the concentration of PINP in serum. After optimizing the conditions, we determined that the best working concentration of the capture antibody and the detection antibody were 3 μg/mL, and the incubation time was 30 minutes. The quantitative assay had a detection range of 5-1 100 ng/mL, with recovery rates between 93%-107% and the minimum detection limit of 1.22 ng/mL achieved. The intra-and inter-assay precisions were lower than 10%. The correlation coefficient of PINP results between this CLIA method and the Roche electrochemiluminescence immunoassay system was 0.906 2. Therefore, this CLIA method is specific and can be used to quantitatively detect the content of PINP in serum, which has the potential to become an auxiliary approach for bone disease examination.
Animals ; Humans ; Immunoassay ; Luminescence ; Mice ; Mice, Inbred BALB C ; Peptide Fragments/isolation & purification* ; Procollagen/isolation & purification*

Objective To establish a rapid diagnosis method for the biological toxicity of soil, accurately and rapidly evaluate the toxicity of contaminated sites and identify the dominant pollutants. Methods Take the soil pollution of a galvanized factory as an example, while the metal concentration level was analyzed and detected, a rapid biological toxicity detection method based on the acute toxicity test of luminescent bacteria （Vibrio qinghaiensis sp.-Q67） was established, and the dominant pollutants were identified by stepwise multiple regression. Results The pollutants came from wastewater and metal plating fragments directly discharged from the manufacturing line of the factory. The concentration of those pollutants was correlated with the acute toxicity of Vibrio qinghaiensis sp.-Q67. The dominant pollutants in the study were zinc （Zn）, aluminum （Al） and copper （Cu）. Conclusion The luminescent bacteria toxicity test method based on Vibrio qinghaiensis sp.-Q67 can conveniently and rapidly assess the degree of toxic damage of polluted soil and identify the dominant pollutants and can be applied to the acute toxicity evaluation of polluted soil.
Luminescence ; Vibrio ; Wastewater

Targeted alpha therapy (TAT) is an active area of drug development as a highly specific and highly potent therapeutic modality that can be applied to many types of late-stage cancers. In order to properly evaluate its safety and efficacy, understanding biokinetics of alpha-emitting radiopharmaceuticals is essential. Quantitative imaging of alpha-emitting radiopharmaceuticals is often possible via imaging of gammas and positrons produced during complex decay chains of these radionuclides. Analysis of the complex decay chains for alpha-emitting radionuclides (Tb-149, At-211, Bi-212 (decayed from Pb-212), Bi-213, Ra-223, Ac- 225, and Th-227) with relevance to imageable signals is attempted in this mini-review article. Gamma camera imaging, single-photon emission computed tomography, positron emission tomography, bremsstrahlung radiation imaging, Cerenkov luminescence imaging, and Compton cameras are briefly discussed as modalities for imaging alpha-emitting radiopharmaceuticals.
Electrons ; Luminescence ; Positron-Emission Tomography ; Radioisotopes ; Radionuclide Imaging ; Radiopharmaceuticals ; Tomography, Emission-Computed ; Tomography, Emission-Computed, Single-Photon

OBJECTIVE: To evaluate the performance of the chemiluminescence immune assay (CLIA) and the electro-chemiluminescence immuneoassay(ECLIA) for Treponemapallidum antibody（anti-TP） screening in blood donors. METHODS: The sero-panel samples from NCCL were tested with ELISA, CLIA and ECLIA assays synchronously to evaluate their performances respectively. RESULTS: The sensitivity and the negative predictive value of the CLIA were 100%, which were the same as one kind of ELISA, and better than the other ELISA; The specificity of the CLIA was 88.46%, the accuracy rate was 97.02%, the positive predictive value was 96.13%, which were higher than both ELISA. Due to the significant interference of sample heat inactivation in ECLIA detection, the result can not demonstrate the true performance of ECLIA in this study. The preliminary result was as follows: the sensitivity was 98.93%, the negative predictive value was 96.75%, and the accuracy rate, specificity and positive predictive value of ECLIA were 97.02%, 91.54% and 97.10% respectively. CONCLUSION Compared with ELISA, the CLIA has higher sensitivity and specificity and can be used for Treponemal antibody screening in blood bank. Unfortunately, the data in this study cannot come to a conclusion for ECLIA and needs more testing.
Blood Donors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Luminescence ; Luminescent Measurements ; Sensitivity and Specificity

OBJECTIVE: To explore the application of blood screening method based on chemiluminescence immunoassay（CLIA）and to evaluate its officacy. METHODS: Screening HBsAg, anti-HCV, anti-HIV and TP was performed on 3,530 voluntary blood donors by ELISA and CLIA, and then all the specimens with ELISA and ELISA/CLIA were further confirmed by NAT; TP single and double positive specimens by ELISA or CLIA were further confirmed by TPPA. RESULTS: The results of CLIA method was well consistent with NAT results, displaying better repeatability and higher sensitivity than ELISA method. For CLIA/ELISA specimens there was a certain false-negative result obtained by ELISA method, especially for blood donors with low virus biter concentration or "window period". CONCLUSION ELISA and CLIA have complementary advantages in blood screening, which can improve the sensitivity of blood screening, reduce the missed detection and shorten detection time. The introduction of CLIA for blood screening is of great importance for ensuring the quality of blood and the safety of clinical transfusion.
Blood Donors ; Hepatitis B Surface Antigens ; Hepatitis B virus ; Luminescence ; Luminescent Measurements ; Mass Screening

PURPOSE: We examined the association between thyroid hormone and lower urinary tract symptoms (LUTS)/benign prostatic hyperplasia (BPH). MATERIALS AND METHODS: A total of 5,708 middle aged men were included. LUTS/BPH were assessed using the international prostate symptom score (IPSS), total prostate volume (TPV), maximal flow rate (Qmax), and a full metabolic workup. Thyroid stimulating hormone (TSH) and free thyroxine (FT4) levels were measured using chemiluminescence immunoassay. We divided participants into quartiles based on their TSH and FT4 levels: first to fourth quartile (Q1–Q4). RESULTS: There was a significant increase in the percentage of men with IPSS>7, Qmax<10 mL/s, and TPV≥30 mL with increase of FT4 quartile. The adjusted odds ratio (OR) for TPV≥30 mL and IPSS>7 were significantly different between FT4 quartile groups (ORs; [5–95 percentile interval], p; TPV≥30 mL, Q1: 0.000 [references]; Q2: 1.140 [0.911–1.361], p=0.291; Q3: 1.260 [1.030–1.541], p=0.025; Q4: 1.367 [1.122–1.665], p=0.002; IPSS>7: Q1: 0.000 [references]; Q2: 0.969 [0.836–1.123], p=0.677; Q3: 1.123 [0.965–1.308], p=0.133; Q4: 1.221 [1.049–1.420], p=0.010). In men with above median levels of testosterone, the FT4 correlated positively with TPV, even after adjusting for confounders. However, the FT4 was not correlated with TPV in men with below median levels of testosterone. TSH was not related to LUTS/BPH measurements. CONCLUSIONS: TPV, IPSS, and Qmax were significantly related to FT4. TPV and IPSS were significantly and independently related to FT4. Additionally, the relationship between FT4 and TPV was distinct when testosterone levels are high.
Humans ; Immunoassay ; Lower Urinary Tract Symptoms ; Luminescence ; Male ; Middle Aged ; Odds Ratio ; Prostate ; Prostatic Hyperplasia ; Testosterone ; Thyroid Gland ; Thyrotropin ; Thyroxine ; Urinary Tract ; Urologic Diseases

BACKGROUND: Both hyperuricemia and hyperhomocysteinemia are known as main risk factors of cardiovascular diseases. There has been, however, no report on the relationship between carotid intima-media thickness (IMT) and homocysteine (Hcy) in hyperuricemic patients. This study aimed to investigate how hyperuricemia is associated with increased carotid IMT with a focus on hyperhomocysteinemia. METHODS: This cross-sectional study included 1,222 patients who visited the Chung-Ang University Hospital Health Promotion Center from January 2013 to December 2015. The serum Hcy levels were estimated with a competitive immunoassay using the direct chemiluminescence method. The carotid IMT was measured by B-mode carotid ultrasonography. The definition of hyperuricemia was a serum uric acid level > 7.0 mg/dL for men or > 5.6 mg/dL for women, and hyperhomocysteinemia was defined as serum levels > 15 μmol/L. RESULTS: The hyperuricemic patients showed significantly higher serum Hcy levels and lower estimated glomerular filtration rate (eGFR) than did normouricemic patients (13.39 ± 4.42 vs. 11.69 ± 3.65 μmol/L, P < 0.001; 85.16 ± 19.18 vs. 96.14 ± 16.63, P < 0.001, respectively). Serum Hcy level (odds ratio [OR], 1.050; 95% confidence interval [CI], 1.009–1.092) and fasting glucose level (OR, 1.018; 95% CI, 1.011–1.026) were independent risk factors for carotid plaque. In patients with hyperuricemia, the serum Hcy levels correlated with the eGFR (γ = −0.478, P < 0.001). The carotid IMT correlated with serum Hcy levels and eGFR (γ = 0.196, P = 0.008; γ = − 0.297, P < 0.001, respectively) but not with the serum lipid profile. CONCLUSION: These results suggest that renal function impairment in hyperuricemic patients may worsen carotid IMT by increasing serum Hcy levels.
Cardiovascular Diseases ; Carotid Intima-Media Thickness ; Cross-Sectional Studies ; Fasting ; Female ; Glomerular Filtration Rate ; Glucose ; Health Promotion ; Homocysteine ; Humans ; Hyperhomocysteinemia ; Hyperuricemia ; Immunoassay ; Luminescence ; Male ; Methods ; Risk Factors ; Ultrasonography ; Uric Acid