Immune Colloidal Gold Technique Research Articles

The effect of carbon nanoparticles vs. immune colloidal gold technique test strips on parathyroid protection in total thyroidectomy: A randomized clinical trial study.

The long-term effect of intraoperative usage of carbon nanoparticles (CN) and parathyroid hormone (PTH) test strip using immune colloidal gold technique (ICGT) is unclear. This study aims to compare the effect of intraoperative usage of CN and ICGT test strips on PG function. This randomized clinical study involved adult patients who underwent total thyroidectomy. They were randomly allocated into three groups (control, CN, and ICGT group). Clinical data were analyzed. Each group involved 98 patients. Serum calcium and PTH concentrations at 24 h postoperatively (PTH24h) were higher in CN group. The parathyroid function recovered quicker in CN group. Use of CN increased in situ PG preservation and PTH24h. Mediation analysis indicated that 23.05% of the total effect of CN on PTH24h was attributed to PGRIS. CN holds promise to improve in situ PG preservation and protect PG vasculature, thereby reducing the incidence of early hypoparathyroidism. The value of ICGT test strips for PG protection is dubious.

Increased clonal dissemination of OXA-232-producing ST15 Klebsiella pneumoniae in Zhejiang, China from 2018 to 2021

BackgroundOXA-232-producing Klebsiella pneumoniae was first identified in China in 2016, and its clonal transmission was reported in 2019. However, there are no prevalence and genotypic surveillance data available for OXA-232 in China. Therefore, we investigated the trends and characteristics of OXA-232 type carbapenemase in Zhejiang Province, China from 2018 to 2021.MethodsA total of 3278 samples from 1666 patients in the intensive care units were collected from hospitals in Zhejiang Province from 2018 to 2021. Carbapenem-resistant isolates were initially selected by China Blue agar plates supplemented with 0.3 μg/ml meropenem, and further analyzed by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry identification, immune colloidal gold technique, conjugation experiment, antimicrobial susceptibility testing and whole genome sequencing.ResultsA total of 79 OXA-producing strains were recovered, with the prevalence increased from 1.8% [95% confidence interval (CI): 0.7–3.7%] in 2018 to 6.0% (95% CI: 4.4–7.9%) in 2021. Seventy-eight strains produced OXA-232 and one produced OXA-181. The blaOXA-232 gene in all strains was located in a 6141-bp ColKP3-type non-conjugative plasmid and the blaOXA-181 gene was located in a 51,391-bp ColKP3/IncX3-type non-conjugative plasmid. The blaOXA-232-producing K. pneumoniae was dominated (75/76) by isolates of sequence type 15 (ST15) that differed by less than 80 SNPs. All OXA-producing strains (100%, 95% CI: 95.4–100.0%) were multidrug-resistant.ConclusionsFrom 2018 to 2021, OXA-232 is the most prevalent OXA-48-like derivative in Zhejiang Province, and ST15 K. pneumoniae isolates belonging to the same clone are the major carriers. The transmission of ColKP3-type plasmid to E. coli highlighted that understanding the transmission mechanism is of great importance to delay or arrest the propagation of OXA-232 to other species.

Evaluation of the immune colloidal gold technique for BP180-NC16A-specific antibodies in the quick diagnosis and monitoring of bullous pemphigoid

BackgroundBullous pemphigoid (BP) mostly involves elderly patients. The diagnosis of BP requires special immunological tests, which makes some patients unable to be diagnosed and treated timely. ObjectiveThe accuracy and application value of immune colloidal gold technique (ICGT) in BP were evaluated. The colloidal gold was conjugated with recombinant BP180 NC16A protein and mouse IgG antibody. As the test and control lines, the mouse-anti-human IgG and goat-anti-mouse IgG, respectively, were blotted on the nitrocellulose membrane. Methods414 serum samples of consecutive patients with suspected BP and 15 samples from healthy donors were recruited. The consistency between ICGT and ELISA, and between serum and plasma/whole blood were evaluated. Subgroup analyses were performed in terms of clinical characteristics. We also followed up 65 BP patients’ strip results to explore the predictive value of ICGT. ResultsStrong agreements between ICGT and ELISA(κ = 0.902) and between plasma/whole blood and serum samples (κ = 0.980) with good stability were observed. The ICGT achieved sensitivity of 93.9%, and specificity of 97.6%. In subgroup analysis, the sensitivity was significantly higher in older patients (96.3%), and with more typical lesions such as blisters (96.2%) and erosions (92.4%). In follow-up, we also found BP patients who kept ICGT-negative in remission state all got consecutive positive strips 1–3 weeks prior to mild new activity or flare. ConclusionICGT shows high potential as a rapid and stable option for the diagnosis and monitoring of BP. Further investigations are needed to re-evaluate this technique in a prospective study with a multicenter design.

Review detection of Newcastle disease virus.

Newcastle disease (ND) is an acute and highly contagious disease caused by the Newcastle disease virus (NDV) infecting poultry, which has caused great harm to the poultry industry around the world. Rapid diagnosis of NDV is important to early treatment and early institution of control measures. In this review, we comprehensively summarize the most recent research into NDV, including historical overview, molecular structure, and infection mechanism. We then focus on detection strategies for NDV, including virus isolation, serological assays (such as hemagglutination and hemagglutination-inhibition tests, enzyme linked immunosorbent assay, reporter virus neutralization test, Immunofluorescence assay, and Immune colloidal gold technique), molecular assays (such as reverse transcription polymerase chain reaction, real-time quantitative PCR, and loop-mediated isothermal amplification) and other assays. The performance of the different serological and molecular biology assays currently available was also analyzed. To conclude, we examine the limitations of currently available strategies for the detection of NDV to lay the groundwork for new detection assays.

Whole-genome analysis and description of an IMP-8-producing Ochrobactrum anthropi

To explore the genomic characterization of an IMP-8-producing Ochrobactrum anthropic and give suggestions for the application of antibiotics. In 2021, the infection caused by CRKP was under control after nearly three months of using CAV, however, carbapenem-resistant O. anthropi isolates were collected from a rectal swab sample of a patient with Lumbar Disc Herniation Postoperative Infection. The rectal swab was then enriched in lysogeny broth overnight and inoculated onto China Blue agar plates containing 0.3µg/mL meropenem. And we investigated the characteristics of this carbapenem-resistant O. anthropi by MALDI-TOF MS, Immune colloidal gold technique, conjugation experiment, whole genome sequencing and antimicrobial susceptibility testing. Antimicrobial susceptibility testing showed that the O. anthropi were resistant to imipenem, cefmetazole, ceftazidime, cefotaxime, piperacillin/tazobactam, sulbactam/cefopcrazone, ceftazidime/avibactam, cefepime, ciprofloxacin, aztreonam, and not susceptible to meropenem, ertapenem, polymyxin B, tigecycline, amikacin. Immune colloidal gold technique reflected that this strain produced IMP carbapenemases, and the presence of IMP-8 was verified by WGS, which was located in a 21,442 bp, nonconjugative plasmid. Improper antibiotic treatment can cause intestinal flora imbalance and even bacteremia in patients, we should use antibiotics wisely and develop individualized treatment options.

Emergence and Genomic Characterization of a KPC-2-, NDM-1-, and IMP-4-Producing Klebsiella michiganensis Isolate.

A rectal swab sample was collected from a patient with Guillain–Barré syndrome and enriched in lysogeny broth. Carbapenem-resistant bacteria were selected by China Blue agar plates containing 0.3 μg/ml meropenem. Carbapenemase-producing Klebsiella michiganensis was identified and characterized by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF), immune colloidal gold technique, a conjugation experiment, PCR analysis, and antimicrobial susceptibility testing. The genome of K. michiganensis was determined by whole genome sequencing. Antimicrobial susceptibility testing showed that the K. michiganensis was resistant to imipenem, meropenem, ertapenem, cefmetazole, ceftazidime, cefotaxime, piperacillin/tazobactam, sulbactam/cefoperazone, ceftazidime/avibactam, cefepime, and aztreonam while susceptible to polymyxin B, ciprofloxacin, tigecycline, and amikacin. Immune colloidal gold technique suggested that this strain co-produced three different carbapenemases [Klebsiella pneumoniae carbapenemase (KPC), New Delhi metallo-beta-lactamase (NDM), and Imipenem (IMP)]. Whole genome sequencing analysis indicated that this strain belonged to ST91, and blaKPC–2, blaNDM–1, and blaIMP–4 were carried on different conjugative plasmids. Besides, the co-existence and transferability of blaKPC–2, blaNDM–1, and blaIMP–4 in K. michiganensis facilitates the potential horizontal dissemination and nosocomial spread of resistance genes among multidrug-resistant organisms.

The kidnapping of mitochondrial function associated with the SARS-CoV-2 infection.

Infection by the Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) leads to multi-organ failure associated with a cytokine storm and septic shock. The virus evades the mitochondrial production of interferons through its N protein and, from that moment on, it hijacks the functions of these organelles. The aim of this study was to show how the virus kidnaps the mitochondrial machinery for its benefit and survival, leading to alterations of serum parameters and to nitrosative stress (NSS). In a prospective cohort of 15 postmortem patients who died from COVID-19, six markers of mitochondrial function (COX II, COX IV, MnSOD, nitrotyrosine, Bcl-2 and caspase-9) were analyzed by the immune colloidal gold technique in samples from the lung, heart, and liver. Biometric laboratory results from these patients showed alterations in hemoglobin, platelets, creatinine, urea nitrogen, glucose, C-reactive protein, albumin, D-dimer, ferritin, fibrinogen, Ca²⁺, K⁺, lactate and troponin. These changes were associated with alterations in the mitochondrial structure and function. The multi-organ dysfunction present in COVID-19 patients may be caused, in part, by damage to the mitochondria that results in an inflammatory state that contributes to NSS, which activates the sepsis cascade and results in increased mortality in COVID-19 patients.

A Rapid Intraoperative Parathyroid Hormone Assay Based on the Immune Colloidal Gold Technique for Parathyroid Identification in Thyroid Surgery.

ObjectiveA novel immunochromatographic test strip method was developed to detect tissue parathyroid hormone (PTH) using the immune colloidal gold technique (ICGT). The accuracy and application value of this method for intraoperative parathyroid identification were evaluated.MethodsSerum samples were collected to measure PTH by both ICGT and electrochemiluminescence immunoassay (ECLIA). Patients who underwent unilateral and total thyroidectomy were enrolled to evaluate the feasibility and clinical efficacy of rapid intraoperative identification of parathyroid glands via PTH determination using ICGT. Two sample preparation methods, fine needle aspiration (FNA) and tissue block homogenate (TBH), were used for PTH-ICGT analysis.ResultsBablok analysis showed a linear relationship between the serum PTH measurements obtained by ICGT and ECLIA. Non-parathyroid tissues had much lower PTH concentrations (14.8 ± 2.1 pg/ml, n = 97) detected by ICGT, compared to the parathyroid gland tissues (955.3 ± 16.1 pg/ml, n = 79; P < 0.0001), With biopsy results as the standard, ICGT showed higher diagnosis rates as compared with direct visual inspection, for identifying both parathyroid glands (97.4 vs. 78.2%) and non-parathyroid tissues (100 vs. 68.9%). The cut-off values for parathyroid identification by FNA and TBH methods were 63.99 and 136.30 pg/ml, respectively. The detection time was 2 min by TBH method for in vitro tissue detection and 6 min by FNA method for in situ tissue detection, both of which were faster than traditional intraoperative cryopathological examination (usually >30 min). Intraoperative application of ICGT method was associated with higher postoperative serum calcium and blood PTH levels at 1 and 3 months as well as a lower incidence of postoperative transient hypocalcemia, as compared with direct visual inspection.ConclusionPTH-ICGT assay shows high potential as a rapid, novel alternative for intraoperative parathyroid identification.

Comparison of different detection methods for Mycoplasma pneumoniae infection in children with community-acquired pneumonia

BackgroundDue to the lack of a sensitive, specific and rapid detection method, aetiological diagnosis of pneumonia caused by Mycoplasma pneumoniae (M. pneumoniae, MP) is a constantly challenging issue. This retrospective study aimed to compare the diagnostic methods for Mycoplasma pneumoniae in children and evaluate their values.MethodsFrom November 2018 to June 2019, 830 children with community-acquired pneumonia were selected from the Department of Respiratory Medicine, Shanghai Children’s Medical Center. On the first day of hospitalization, sputum, throat swab and venous blood samples were collected to analyse MP-IgM (particle agglutination, PA), MP-IgM (immune colloidal gold technique, GICT), MP-DNA, MP-RNA (simultaneous amplification and testing, SAT) and MP-DNA (real-time polymerase chain reaction, RT-PCR).ResultsAmong these 830 children, RT-PCR showed that the positive rate was 36.6% (304/830), in which the positive rate of macrolide resistance (A2063G mutation) accounted for 86.2% of cases (262/304). Using RT-PCR as the standard, MP-RNA (SAT) had the highest specificity (97.5%), and MP-IgM (PA) had the highest sensitivity (74.0%) and Youden index (53.7%). If MP-RNA (SAT) was combined with MP-IgM (PA), its Kappa value (0.602), sensitivity (84.2%), specificity (78.7%) and Youden index (62.9%) were higher than those of single M. pneumoniae detection.ConclusionsOur research indicated that a combination of MP-RNA (SAT) plus MP-IgM (PA) might lead to reliable results as an early diagnostic method for children with clinical manifestations of Mycoplasma pneumoniae pneumonia.

Identification of Mycobacteria isolated from AIDS patients

Obstract: To identify Mycobacteria isolated from AIDS patients and to evaluate the diagnostic value of MBP64 antigen Colloidal Gold Diagnostic Kit in distinguishing Mycobacterium tuberculosis(MTB) and Nontuberculous mycobacteria (NTM). Methods: A total of 140 Mycobacteria isolates from AIDS patients were collected and identified by using 16SrDNA PCR assay as well as immune colloidal gold technique (MBP64 antigen Colloidal Gold Diagnostic Kit). The sensitivity and specificity of Colloidal Gold method was analyzed. Results: With the results of 16SrDNA PCR assay as gold standard, the sensitivity and specificity of Colloidal Gold method in distinguishing MTB and NTM were 96.00% and 96.84%, respectively. Among 140 samples, 114 were identified by 16SrDNA PCR assay, among which 50 (43.86%) were identified as MTB, and 63 (55.26%) were identified as NTM. The most popular NTM in AIDS patients were Mycobacterium gordonae, Mycobacterium avium and Mycobacterium kansasii. Conclusion: NTM is of high prevalence in AIDS patients, and MBP64 antigen Colloidal Gold Diagnostic Kit can be used as an effective technique in distinguishing NTM and MTB.

Screening of anti-M2 mitochondrial antibody in elderly population

Serum anti-M2 mitochondrial antibody (AMA-M2) was screened with immune colloidal gold technique in 6 008 individuals aged over 60 years who underwent health check-up in Xuhui District of Shanghai, the positive samples were retested with ELISA method. The results showed that 1.31%(79/6 008) individuals were AMA-M2 positive; the positive rate was 0.82% (18/2 186) in males and 1.60% (61/3 822) in females. Eleven subjects with positive AMA-M2 underwent further investigation, 8 cases were diagnosed as primary biliary cirrhosis (PBC) and the remaining 3 were highly suspected as PBC. All 11 individuals were abnormal in biochemical tests of liver function and/or clinical symptoms. Key words: Liver cirrhosis, biliary; Mass screening

Identification and characterization of a novel gene, c1orf109, encoding a CK2 substrate that is involved in cancer cell proliferation

BackgroundIn the present study we identified a novel gene, Homo Sapiens Chromosome 1 ORF109 (c1orf109, GenBank ID: NM_017850.1), which encodes a substrate of CK2. We analyzed the regulation mode of the gene, the expression pattern and subcellular localization of the predicted protein in the cell, and its role involving in cell proliferation and cell cycle control.MethodsDual-luciferase reporter assay, chromatin immunoprecipitation and EMSA were used to analysis the basal transcriptional requirements of the predicted promoter regions. C1ORF109 expression was assessed by western blot analysis. The subcellular localization of C1ORF109 was detected by immunofluorescence and immune colloidal gold technique. Cell proliferation was evaluated using MTT assay and colony-forming assay.ResultsWe found that two cis-acting elements within the crucial region of the c1orf109 promoter, one TATA box and one CAAT box, are required for maximal transcription of the c1orf109 gene. The 5′ flanking region of the c1orf109 gene could bind specific transcription factors and Sp1 may be one of them. Employing western blot analysis, we detected upregulated expression of c1orf109 in multiple cancer cell lines. The protein C1ORF109 was mainly located in the nucleus and cytoplasm. Moreover, we also found that C1ORF109 was a phosphoprotein in vivo and could be phosphorylated by the protein kinase CK2 in vitro. Exogenous expression of C1ORF109 in breast cancer Hs578T cells induced an increase in colony number and cell proliferation. A concomitant rise in levels of PCNA (proliferating cell nuclear antigen) and cyclinD1 expression was observed. Meanwhile, knockdown of c1orf109 by siRNA in breast cancer MDA-MB-231 cells confirmed the role of c1orf109 in proliferation.ConclusionsTaken together, our findings suggest that C1ORF109 may be the downstream target of protein kinase CK2 and involved in the regulation of cancer cell proliferation.

Fibronectin EIIIA splicing variant: A useful contribution to forensic wounding interval estimation

A fibronectin splicing variant, Fn containing an extra type III domain (EIIIA in rat, ED1 or EDA in human) has recently attracted more attention because of its sensitive response in injury of adult tissue. The characteristic that this form is absent in adult tissue while it is commonly expressed in fetal tissues and injured adult tissue is useful in estimation of injury interval in forensic science. The regulation of fibronectin splicing was studied by immunohistochemistry by using monoclonal antibody to EIIIA on sections from paraffin embedded rat skin sample after contusion. The results indicated that epidermic cells and hair follicle epithelium of rat skin were found positive staining at 6 h after injury, and the color became darker with prolonged wounding time. Based on the above result, we invented a type of test paper assayed for EIIIA-Fn splicing variant by using immune colloidal gold technique. After dipping one end of test paper in the supernatant fluid of tissue homogenates of injury skin for few minutes, detecting line could be found. It showed that all experimental injured samples revealed positive staining; the darkness of positive staining was dependent on the injury time, while control normal skin cannot found positive staining. It is concluded that there is a close relationship between the expression of EIIIA-Fn splicing variant and wounding interval, and that the gold-labeled test paper can be useful in distinguishing ante- and post-mortem injury, and in estimation of wounding interval in forensic science.