To explore the application effect of rare phenotype of common pathogenic bacteria in clinical microbiological examination practice.

The knowledge related to the rare phenotype of common pathogenic bacteria was summarized and introduced into the teaching process of clinical microbiology laboratory practice.

The rare phenotype of common pathogenic bacteria enriches the teaching content, arouses students' initiative and enthusiasm, and improves the quality and effect of clinical microbiological examination practice teaching.

It is an effective teaching method to introduce the rare phenotype of common pathogenic bacteria into the practice teaching of clinical microbiological examination.

To establish an efficient, sensitive and accurate method for the detection of dermatophytosis by using the real-time fluorescence technique based on loop-mediated isothermal amplification, so as to realize the rapid laboratory diagnosis of dermatophytosis.

By searching and downloading the rRNA sequences of the strains with high clinical isolation rates of Trichophyton, Microsporum and Epidermophyfon from the NCBI website, DNASTAR Lasergene software was used for analysis and comparison to find the highly conserved target sequences of Trichophyton, which were different from those of other genera. LAMP primers that included a pair of internal primers, external primers and ring primers were designed using Primer Explorer V5. The NBRC6202 genomic DNA of Trichophyton mentagrophytes was extracted as a template for screening primers and a LAMP reaction system was established for the detection of Trichophyton. The sensitivity and stability of this reaction system were tested by using the ATCC28188 genomic DNA of T. rubrum. The specificity of LAMP reaction system was verified by the genomic DNA of other fungi and bacteria other than Trichophyton.

Among the 4 sets of primers screened, primer2 had a good amplification effect and was able to detect T. mentagrophytes within 22 min. Moreover, T. rubrum, T. interdigitale, T. verrucosum, T. schoenleinii and T. violaceum could be specifically detected. The minimum detection limit of LAMP detection system established in this study was 1 pg/μL. The average CT value and CV% were 9.24±0.30, 3.29%, 10.57±0.54, 5.12%, 15.95±0.52, 3.24% at 10 000 pg/μL, 100 pg/μL, and 1 pg/μL, respectively. The peaking time was stable.

The LAMP method of Trichophyton showed superior performance in specificity, sensitivity and stability, which can quickly and accurately detect common strains of Trichophyton, and had a good clinical application prospect.

To explore the expression characteristic of serum melanoma cell adhesion molecule (MCAM) in patients with hepatocellular carcinoma (HCC), evaluate the clinical significance of candidate MCAM in thediagnosis of HCC.

Eighty-three HCC patients and eighty-five healthy individuals at Shanghai Ruijin Hospital and Shanghai Tenth People’s Hospital from November 2013 to February 2015 were considered eligible for this study. Serum MCAM and cluster of differentiation (CD)₁₆₆ were examined using enzyme-linked immunosorben assay(ELISA). Analysis the relationship between MCAM and those seven indexes including alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyl transpeptidase (γ-GT), total bileacid (TBA), total protein (TP) and alpha fetal protein (AFP) by using spearman. Meanwhile, to evaluate the specificity and sensitivity of MCAM in the diagnosisof HCC through the receiver-operating characteristics curve.

Serum MCAM, serum CD₁₆₆, AFP of healthy individuals were (2 776.7±25.8) ng/L, (222.6±74.9) ng/L, (21.7±9.6) μg/L and those of patients with HCC were (20 453.3±110.2) ng/L, (5 702.0±99.0) ng/L, (275.7±83.4) μg/L. A positivecorrelation was found between serum MCAM and CD₁₆₆(r=0.875, P<0.001). Besides, a positivecorrelation was also found between serum MCAM and AFP(r=0.858, P<0.001). In patients with HCC, the data of γ-GT, TBA, ALP, ALT, AST and TP were collected as follows: (120.8±10.0) IU/L, (123.6±24.8) μmol/L, (158.2±15.4) IU/L, (69.3±8.3) IU/L, (135.7±12.8) IU/L, (67.3±8.7) μg/L. Serum MCAM was also positively correlated with CD₁₆₆, ALT, AST, ALP, γ-GT TBA(r of 0875, 0.407, 0.831, 0.456, 0.670, 0.685, P<0.001). The areaunder the receiver operating characteristic curve for serum-MCAM was 0.940, for the differentiation of HCC patients from healthy individualswith a cut-off of 2 397.6 μg/L, sensitivity 90.4% (152/168), specificity 94.1%(158/168). AUC-ROC also indicated that the combination of MCAM and CD₁₆₆ with AFP(0.984, 95%CI 0.969-1.000), sensitivity 98.8% (160/168), specificity 94.1% (158/168) to diagnose HCC was only slightly better than AFP (0.931, 95%CI 0.887-0.974), sensitivity 89.2% (150/168), specificity 94.1% (158/168) or MCAM (0.940(95% CI 0.900-0.981), sensitivity 90.4% (152/168), specificity 94.1% (158/168) alone or the combination of MCAM and CD₁₆₆ (0.976, 95% CI 0.955-0.998), sensitivity 97.6% (164/168), specificity 92.9% (156/168).

Serum MCAM is a novel diagnostic tumor marker for HCC. With high sensitivity and specificity, MCAM could have a good diagnostic efficiency as a novel hepatocellular carcinoma tumor marker in the near future. But this still need to expand the sample size to further verification. The single examination of AFP, MCAM or the combination of MCAM with CD₁₆₆ are all hetpful for diagnosis of HCC in clinical application.