Association analysis of tissue inhibitor of metalloproteinase2 gene polymorphisms with COPD in Egyptians.

Proteinase/antiproteinase imbalance is recognized to play an important role in the pathogenesis of chronic obstructive pulmonary disease (COPD). A relative increase in the activities of matrix metalloproteinases might be caused by mutations of tissue inhibitor of metalloproteinase2 (TIMP2). Recently, two polymorphisms of the TIMP2 gene, +853 G/A and -418 G/C (+551 and -720 from the translation initiation site), have been shown to be associated with the development of COPD in the Japanese population. In this study, a case-control association analysis for these polymorphisms was conducted in the Egyptian population using 106 COPD patients and 72 healthy controls. The genotype frequency of +853 G/A was significantly different between the patient and the control groups (P = 0.029), although no significant difference was detected in the allele frequency between the two groups. These results suggest that the +853 G/A polymorphism of the TIMP2 gene might be associated with COPD across ethnicities. In contrast, neither the distributions of genotype nor allele frequencies of -418 G/C were significantly different between the two groups, raising the possibility that a combination of different genetic factors contributes to the development of COPD in different ethnic groups.

Rapid diagnosis of adenoviral conjunctivitis on conjunctival swabs by 10-minute immunochromatography.

PURPOSE: Several methods are available for the diagnosis of acute conjunctivitis, all of which are time-consuming or require the use of a well-equipped laboratory. A new method, immunochromatography (IC), for detecting the presence of adenovirus (Ad) has been developed. Two direct rapid tests to detect Ad antigen, IC and enzyme immunoassay (EIA), were compared with regard to sensitivity, specificity, and technical complexity. METHODS: The study materials consisted of 130 swabs from patients with conjunctivitis (95 samples of adenoviral conjunctivitis proven by positive virus DNA on polymerase chain reaction [PCR], 35 samples of nonadenoviral conjunctivitis proven by PCR). IC is a one-step procedure that detects the presence of adenoviral antigen by sandwich EIA on a paper disc. RESULTS: In 95 adenoviral DNA-positive samples by PCR, the sensitivity and specificity of IC were 54.7% and 97.1%, respectively, whereas those of EIA were 50.5% and 100%, respectively. By IC, PCR-positive Ad type 3 was recognized in 31%; Ad4 in 100%; Ad7 in 60%; Ad8 in 67%; and Ad37 in 59%, showing similar positivity rates for different serotypes (except Ad7) to those using EIA. Visual determination of the presence of Ad took an average of 10 minutes by IC compared with 70 minutes by EIA. CONCLUSIONS: These results indicate that IC is a more rapid and easier test compared with EIA, and it has high specificity. Detection of Ad antigen by this simple and rapid method will serve physicians as a useful tool for early diagnosis and prevention of adenoviral conjunctivitis.

[Rapid detection and identification of human adenovirus directly from conjunctival scrapings by polymerase chain reaction and restriction fragment length polymorphism analysis].

Polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analysis were combined for detection and identification of adenovirus (Ad), a common agent of conjunctivitis in Japan. Nested-PCR with two primer sets that hybridize to the conserved region for hexon protein of 14 prototypes of Ad serotype 1 to 8, 11, 14, 19, 37, 40, and 41, amplified 956 bps DNA fragment. The amplified fragments from 14 prototypes were completely differentiated with the combination of three restriction endonucleases, Eco T14I, Hae III, and Hin fI. We applied this new method to 70 conjunctival scrapings from patients with conjunctivitis, and compared the results with those of the combination of culture isolation and neutralization test. PCR was positive in 38 out of 70 samples (54.3%), whereas 33 of 70 samples (47.1%) were positive by cell culture. Compared with cell culture isolation, the PCR method had a sensitivity of 100% (33 of 33). Positive PCR samples were further classified into Ad 37 (44.7%), 3 (39.5%), 11 (7.9%), 8 (5.3%), and 4 (2.6%) by PCR-RFLP analysis. Of five samples that were PCR positive and cell culture negative, three samples were Ad 37 and two were Ad 8 by PCR-RFLP analysis. These differentiations of cell culture positive samples were identical to the results of the neutralization test. It took only about three days to detect and identify Ad by PCR-RFLP analysis, whereas it took at least two weeks by culture isolation and neutralization test. Our newly developed method of detecting and typing human Ad by PCR-RFLP analysis is more sensitive, accurate, and prompt than the conventional cell culture isolation and neutralization test.