Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hypothetical Protein Gene in Escherichia Coli Clinical Isolates.

BACKGROUND: Escherichia coli is the most common pathogenic bacteria that frequently causes life-threatening opportunistic human infections, diarrhea, and septicemia in immunocompromised hosts. METHODS: This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of a hypothetical protein from an E. coli-specific gene (GenBank ID: 13702648). The gene was obtained through local and online BLAST, and specific primers were designed for this gene. Reaction conditions were optimized at 65ºC for 30 minutes and 80ºC for 2 minutes, whereas the reaction system contained 5.2 mM Mg2+, 8 U of Bst 2.0 DNA polymerase, 1.4 mM deoxyribonucleotide, and 0.2 and 1.6 µM of the outer and inner primers, respectively. The LAMP method was evaluated using 240 strains of E. coli and 150 strains of non-E. coli. RESULTS: Positive reactions were observed on all 240 strains of E. coli while all non-E. coli strains were negative. Plasmids with the specific gene and mice blood with E. coli were used for sensitivity analysis. The detection limit of LAMP was 100 bacterium/reaction. CONCLUSIONS: Results showed that the LAMP targeted to the hypothetical protein (GenBank ID: 13702648) is a fast, specific, sensitive, inexpensive, and suitable method for the detection of E. coli.

[Expression of MICA/B protein in esophageal cancer and its clinical significance].

OBJECTIVE: To explore the expression of MICA/B in human esophageal cancer, and to analyze its correlation with clinicopathological features. METHODS: The expression of MICA/B in 40 cases of esophagus carcinoma and corresponding normal esophageal mucosa tissues were examined by immunohistochemistry. RESULTS: The positive rate of expression of MICA/B protein in the esophageal carcinoma was 75.0% (30/40), and that in the corresponding normal esophageal mucosa was 0 (0/40). Up-regulation of MICA/B expression was found in the esophageal carcinomas. The expression of MICA/B was related with histological grade of the esophageal carcinoma (P = 0.012). CONCLUSION: MICA/B protein plays an important role in the esophageal carcinogenesis, and my become a useful molecular marker for the diagnosis of esophageal carcinoma.

Analysis of clinical characteristics and differential diagnosis of the lung biopsy specimens in 99 adenocarcinoma cases and 111 squamous cell carcinoma cases: utility of an immunohistochemical panel containing CK5/6, CK34ßE12, p63, CK7 and TTF-1.

This study aimed at evaluating the utility of a panel of antibodies, consisting of cytokeratins (CK) 5/6, CK34ßE12, p63, thyroid transcription factor-1 (TTF-1), and CK7 for distinguishing between squamous cell carcinoma (SCC) and adenocarcinomas (Ad), as well as their expression with clinicopathological parameters and prognosis in SCC and Ad. 111 SCC of small biopsy specimens and 99 cases of Ad were stained by immunohistochemistry, among which 76 SCC and 64 Ad had complete follow-up data. Most of SCC displayed CK5/6 (91/99, 91.92%), CK34ßE12 (83/99, 83.84%), p63 (96/99, 96.97%), and most of Ad showed expression of CK7 (89/111, 80.18%) and TTF-1 (105/111, 94.59%). The combination of CK5/6/CK34ßE12/p63 seems to be useful for differentiating SCC from Ad with 100% sensitivity and 97.30% specificity, and TTF-1 was a useful biomarker for Ad with 94.59% sensitivity and 100% specificity. There were differences between CK5/6, p63, and TTF-1 expression with tumor differentiation (p<0.05) in SCC or Ad. Univariate analysis indicated that patients with high TTF-1 expression predicted better prognosis in Ad patients. Multivariate analysis showed that TTF-1 expression (HR=0.340, 95% CI 0.143-0.811, p=0.015) was a good independent predictor of Ad patient survival.