Quick and accurate genotyping of hepatitis C virus (HCV) is becoming increasingly important for clinical management of chronic infection and as an epidemiological marker. Furthermore, the incidence of HCV infection with mixed genotypes has clinical significance that is not addressed by most genotyping methods. We have developed a fluorescence-based genotyping assay called primer-specific extension analysis (PSEA) for the most prevalent HCV genotypes and have demonstrated the capacity of PSEA-HCV for detecting mixedgenotype HCV infections. PSEA-HCV detects genotype-specific sequence differences in the 5 untranslated region of HCV in products amplified by the COBAS AMPLICOR HCV Test, v2.0. Simulated mixed HCV infection of plasma with RNase-resistant RNA controls demonstrates that PSEA-HCV can detect as many as five genotypes in one specimen. Furthermore, in dual-genotype simulations, PSEA-HCV can unequivocally detect both genotypes, with one genotype representing only 3.1% of the mixture (313/10,000 IU in starting plasma). Compared to INNO-LiPA HCV II, both assays determined the same genotype for 191/199 (96%) patient specimens (175 subtype and 16 genotype-only identifications). Following the initial evaluation, PSEA-HCV was used routinely to genotype HCV from patient specimens submitted to our laboratory (n ‫؍‬ 312). Seventeen (5.4%) mixed infections were identified. The distribution of single-infection HCV genotypes in our population was 60.9% type 1 (n ‫؍‬ 190), 12.8% type 2 (n ‫؍‬ 40), 20.2% type 3 (n ‫؍‬ 63), 0.3% type 4 (n ‫؍‬ 1), and 0.3% other (n ‫؍‬ 1). In conclusion, PSEA-HCV provides an inexpensive, high-throughput screening tool for rapid genotyping of HCV while reliably identifying mixed HCV infections.Infection by hepatitis C virus (HCV) is the leading cause of chronic liver disease worldwide (19). The overall prevalence of HCV infection in the United States is 1.8%, with most of the patients unaware of their infection and at risk for developing cirrhosis and hepatocellular carcinoma (15,21). HCV is a positive-sense, single-stranded RNA virus that displays extensive genetic heterogeneity (1). At least six major HCV genotypes comprising numerous, more closely related subtypes have been identified (26). HCV genotypes display significant differences in their global distribution and prevalence, making genotyping a useful method for determining the source of HCV transmission in an infected localized population (11). Furthermore, in addition to viral load and liver histology, the genotype of the infecting HCV strain appears to be an important determinant of the severity and aggressiveness of liver infection, as well as patient response to antiviral therapy (26). Consequently, several methods for genotyping HCV have been developed, including direct DNA sequencing (2, 5, 6), typespecific PCR (17), restriction fragment length polymorphism (16), line probe assays (22, 23), primer-specific and mispair extension analysis (7,8), heteroduplex mobility analysis by temperature gradient capillary electrophoresis (14)...

The case of a 76-year-old white woman with squamous cell carcinoma of the thyroid gland arising in a background of Hashimoto's thyroiditis is presented. Squamous cell carcinoma of the thyroid gland is a very rare and aggressive tumor, and only a few cases are reported as arising in Hashimoto's thyroiditis. Although Hashimoto's thyroiditis alone does not necessarily predispose patients to malignancy, the finding of a prominent nodule in such a patient should suggest the possibility of malignancy, and an aggressive search and treatment regimen, including surgical intervention, should be pursued.

We used PCR for hepatitis C virus (HCV) genotyping with type-specific primers from the core and NS5 genes. Type I was predominant in the general population (58% in blood donors) as well as in different risk groups, such as intravenous drug abusers (58%), blood transfusion recipients (64%), hemophiliacs (62%), and patients with HCV chronic liver disease (76%). Types 1, 111, and IV were less prevalent in Canada, being found in 10.92, 6.72, and 5.88% of the population, respectively. The type II core primer was not type specific and reacted with the majority of our type I HCV samples, suggesting a false-positive dual infection with two

HCV infections in hemophiliacs may be less severe than in HCV infected patients without hemophilia.