Hybrid chain reaction and selective recognition-based homogeneous dual-fluorescence analysis of circulating tumor cells in clinical ovarian cancer samples.

BACKGROUND: Oncological analysis is important in tumor diagnosis. We constructed a dual-fluorescence and binary visual analysis system for circulating tumor cells (CTCs) using the folate receptor as a biomarker, combined with hybridization chain reaction and nanomaterial amplification. This strategy integrates terminal protection, selective recognition properties of N-methyl mesoporphyrin IX and CdTe quantum dots for Cu2+ and double-stranded templated copper nanoparticles, and inkjet printing technology. RESULTS: In fluorescence mode, folate receptor and A2780 ovarian cancer cells were specifically detected with a limit of detection of 0.1 fg mL-1, and 10 cells mL-1 were observed. The detection limits of both the color and distance reading modes were comparable to those obtained in fluorescence mode. The applicability of the method for quantifying CTCs was validated using 27 (6 negative and 21 positive) clinical ovarian cancer samples; the results agreed with those of both the clinical folate receptor-polymerase chain reaction kit and radiological and pathological results. SIGNIFICANCE: This dual-fluorescence and binary visual CTCs detection method provides multiple options for clinical tumor liquid biopsy.

[Preliminary Study on PCR Method for Laboratory Diagnosis with Fecal Specimens of Entamoeba histolytica Infection].

OBJECTIVE: To develop a PCR method for Entamoeba histolytica（ E.histolytica) detection in fecal specimens, and to compare the performance of PCR to that of microscopy and ELISA. METHODS: Two pairs of self-designed primers and 2 pairs of primers from references based on small subunit ribosome RNA (SSU rRNA) fragment of E. histolytica standard strain were synthetized. DNA from E. histolytica reference strain were amilified by the conventional PCR using the 4 pairs of primers. 221 stool samples from diarrhea patients were collected and detected for E. histolytica by three methods: Entamoeba trophozoites and cysts detection by microscopy, E. histolytica-specific antigen detection using enzyme-linked immunosorbent assay (ELISA) kit ( E. HISTOLYTICA II), amplification of SSU rRNA fragment of E. histolytica by PCR method. Positive rate of three methods were compared by chi-square test, and Kappa test was applied to determine the concordance among the three methods. RESULTS: Specific fragments of E. histolytica were amplified by the PCR method we developed in this study. Positive rates of PCR, microscopy and ELISA were 2.26%, 0.90% and 9.50%, respectively. The positive rates of the three methods were significantly different ( χ 2 =23.34, P<0.01). The Kappa value of PCR and microscopy was 0.216, and that of PCR and ELISA method was -0.134, both of which showed a weak consistency. PCR results showed best consistency with clinical diagnosis. CONCLUSION: The PCR method we established in this study has a better performance in accuracy than microscopy and ELISA have in laboratory diagnosis of E. histolytica infection.

Anti-centrosome antibodies: Prevalence and disease association in Chinese population.

Anti-centrosome antibodies are rare findings with undefined clinical significance in clinical research. We aimed at investigating the prevalence and clinical significance of anti-centrosome antibodies in Chinese population. Testing results of total of 281,230 ANA-positive sera were retrospectively obtained from West China Hospital Sichuan University in China between 2008 and 2017. We retrospectively collected and analysed the clinical and laboratory data of the patients with positive anti-centrosome antibody. Of the 356 453 patients tested, 281 230 patients had positive antinuclear antibodies (ANAs, 78.9%), but only 78 patients with positive anti-centrosome antibodies (0.022%), of which 74.4% are females. Diagnoses were established in 69 of 78 patients: 37 cases were autoimmune diseases, mainly including undifferentiated connective tissue diseases (UCTD, 9/37), rheumatoid arthritis (RA, 6/37), Sjögren's syndrome (SS, 5/37) and primary biliary cirrhosis (PBC, 5/37), and the remaining were other autoimmune conditions. The most frequent clinical symptoms of the anti-centrosome-positive patients were arthralgia and eyes and mouth drying. Additionally, 86.7% of anti-centrosome antibodies were not associated with other ANA profiles; however, when associated, the most frequent ANA was anti-U1RNP. Anti-centrosome antibodies are featured by a low prevalence and female gender predominance. They are correlated with some specific diseases, both autoimmune diseases, especially UCTD, RA, SS and PBC, and non-autoimmune diseases, such as infection and cancer, which suggests that they might be potential supporting serological markers of these diseases.

Autoimmune disease associated IFIH1 single nucleotide polymorphism related with IL-18 serum levels in Chinese systemic lupus erythematosus patients.

Systemic lupus erythematosus (SLE) has heterogeneous clinical manifestations. IFIH1 (interferon induced with helicase C domain 1) as one of antiviral helicase genes mediating type I interferon production, plays an essential role in the pathogenesis of SLE. The gene variants in IFIH1 could abnormally activate antiviral defenses and increased type I interferon signaling. The present study aimed to validate associations between single nucleotide polymorphisms (SNP) in IFIH1 and the pathogenesis of SLE. In total, rs1990760, rs3747517 and rs10930046 in IFIH1 are genotyped in 400 SLE patients and 659 health controls in Chinese cohort by an improved multiplex ligation detection reaction (iMLDR) technique. Significant associations were observed between alleles of IFIH1 (rs1990760 C > T, P = 0.005, OR = 1.36, 95%CI = 1.10-1.69; rs3747517 T > C, P = 0.004, OR = 1.31, 95%CI = 1.09-1.58, respectively) and SLE susceptibility. IFIH1 rs1990760 TT genotype carriers had lower serum levels of IL-18 (P < 0.001) and granzyme B (P < 0.001) than CC and CT genotype carriers. IFIH1 rs1990760 CT genotype carriers had higher anti-dsDNA-positive than CC and TT genotype carriers. In conclusion, IFIH1 polymorphisms (rs1990760 and rs3747517) were associated with SLE susceptibility and rs1990760 risk T allele related with IL-18 and granzyme B serum levels in SLE patients.