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The World Health Organization (WHO) International Standard (IS) for human immunodeficiency virus type 1 (HIV-1) RNA is only available in limited amounts. It is critical to use the most common HIV-1 genotype as source for secondary standards, e.g. a National Standard for Taiwan. The objective of this study was to establish the first National Standard for HIV-1 RNA NAT assays in Taiwan. A collaborative study, including eleven laboratories from five different countries, was carried out to establish the HIV-1 RNA National Standard by calibration, in International Units (IU), against the WHO HIV-1 RNA IS. The HIV-1 RNA content for the candidate was quantitated by each laboratory in three independent assays and the results were collected and analyzed statistically. Overall, a high level of agreement among results was achieved from different laboratories. In addition, the stability study indicated that the candidate was stable for 24 months at -80±5°C. In conclusion, the candidate standard was established as the first National Standard for HIV-1 RNA for use in NAT assays in Taiwan. The standard is intended to be used for the quality control of HIV-1 NAT assays and as a quantitative reference material for HIV-1 NAT assays., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Intravenous immunoglobulin (IVIG) manufactured from human plasma contains IgG as the primary ingredient, and is used for indications such as immunodeficiency syndrome. Available IVIGs in Taiwan are either manufactured from Taiwanese or North American plasma. The effectiveness of the national immunization program of Taiwan can be evaluated by analyzing and comparing IVIG antibody titers that are induced through the corresponding vaccines (tetanus, diphtheria, and pertussis, measles, rubella, hepatitis A, hepatitis B and varicella). Both enzyme-linked immunosorbent assay (ELISA) and the in vitro neutralization test demonstrated that all IVIGs provide adequate clinical protection against diphtheria and tetanus toxins. ELISA results further revealed that plasma of Taiwanese subjects contains higher levels of pertussis toxin and filamentous hemagglutinin antibodies, when compared to foreign IVIGs. This may be related to the later adoption of acellular pertussis vaccine in Taiwan. Antibodies titers against measles, rubella, hepatitis A, and varicella-zoster virus were otherwise low. Low titers of hepatitis B surface antigen antibodies are present in Taiwanese plasma IVIG, indicating immune memory decline or loss. In conclusion, our results show that Taiwanese IVIG contains varying titers of vaccine-induced antibodies, and serves as a guide for future amendments to Taiwan's immunization program., (Copyright © 2013 The International Alliance for Biological Standardization. Published by Elsevier Ltd. All rights reserved.)

Total mercury concentrations in 69 red mould rice (Monascus) food samples purchased in Taipei, Taiwan, were determined using the direct mercury analyser (DMA) method. The highest mean levels of total mercury in these samples were observed in samples of canned roast eel (20 µg kg(-1)), soy fibrous product (14 µg kg(-1)), red mould rice (8.5 µg kg(-1)), cereal (7.6 µg kg(-1)), dried pork fibre (7.5 µg kg(-1)), and dietary supplement (7.2 µg kg(-1)). All samples analysed had mercury levels below the limit of 50 µg kg(-1) for food standards in rice, edible oil, and fat of Taiwan. The mercury concentration in the Monascus foods differed widely between different food varieties, suggesting that external contamination and raw materials are the main sources of mercury. This study shows that DMA is a simple method proposed for the determination of total mercury in foods containing Monascus. The method requires no sample pre-treatment, and it minimizes potential sources of contamination. The data (42±2 µg kg(-1)) obtained from five analyses of a standard reference material (apple leaves, NIST-1515) showed good agreement with the certified reference value (44±4 µg kg(-1)) provided by the US National Institute of Standards and Technology (NIST). The precision based on the analysis of standard reference material was 4.1%; the average recovery was 95%.

The concentrations of seven trace elements (As, Cd, Cr, Pb, Se, Cu and Zn) in 93 red mould rice (Monascus) food samples in Taipei, Taiwan, were determined by inductively coupled plasma-mass spectrometry (ICP-MS) after wet digestion. The results, calculated in mg kg(-1) (wet weight) for each sample, revealed the general scenario of food safety in Taiwan: As (0.005-12.04), Cd (<0.0005-2.22), Cr (0.014-6.95), Cu (0.012-8.70), Pb (0.001-0.64), Se (<0.001-1.29) and Zn (0.020-67.02). Three food samples were identified with As concentrations higher than regulatory limits: a dietary supplement sample and a seaweed sample with As concentrations that exceeded the limit of Taiwan's health food standard of 2 mg kg(-1), and a canned eel sample with an As concentration that exceeded the limit of Canada's fish standard of 3.5 mg kg(-1). This study suggests that the estimated intakes of these seven trace elements from the consumption of foods containing Monascus pose little risk, as the trace element contents in the majority of samples were lower than the permissible/tolerable intakes per week according to the guidelines recommended by the Food and Agricultural Organization/World Health Organization (FAO/WHO). Moreover, their concentrations in foods containing Monascus differ widely for different food varieties, suggesting that external contaminants and raw materials are the main sources of trace elements. This study shows that ICP-MS is a simple method proposed for the determination of As, Cd, Cr, Pb, Se, Cu, and Zn in foods containing Monascus.

Vibrio parahaemolyticus is an important foodborne pathogen in Taiwan and many other Asian countries. A total of 371 isolates of V. parahaemolyticus collected from patients involved in foodborne illness outbreaks in Taiwan from 1992 to 1995 were characterized. These isolates had typical biochemical characteristics and only 4% were urease positive. The most frequently isolated serovars were O5:K15 (18.5%), O4:K8 (16.2%), O3:K29 (12.5%), O1:K56 (8.3%), O2:K3 (6.5%), and O4:K12 (6.0%). Most of the isolates were susceptible to nalidixic acid, tetracycline, tobramycin, cephalothin, and gentamicin. About 10% of the isolates were resistant to seven or more antibiotics. Approximately 92.4% of these V. parahaemolyticus showed beta-hemolysis on Wagatsuma blood agar plate and approximately 62.1% of these isolates exhibited detectable amounts of thermostable direct hemolysin. Most of the isolates examined exhibited two copies of tdh genes on the 1.3- and 2.5-kb HindIII-digested chromosome fragments with several variations on other fragments. A pulsed-field gel electrophoresis (PFGE) subspecies typing scheme was used to analyze these domestic isolates and the O3:K6 strains from Japan, Korea, and Taiwan. Fifty seven patterns were differentiated with A, B, C, E, and H being the major domestic types (cumulatively 76% of isolates), while O3:K6 strains (PFGE type I), abruptly occurring since 1996, were genetically distant from the major domestic types.

A total of 95 chicken samples that consisted of 34 whole chickens, 32 organs (gizzards and livers), and 29 chicken parts (drumsticks, wings, and breasts), collected from traditional retail markets (no chilling facilities) and supermarkets in Taipei, were examined for the occurrence of enteropathogenic campylobacters. Three selective media, Peterz's charcoal cefoperazone deoxycholate agar, Campy-Cefex agar, and charcoal-based selective medium, were evaluated for their efficacy to isolate Campylobacter spp. from chicken samples. The results showed that there were no differences among the three media to isolate Campylobacter spp. from all chicken samples (P > 0.05). However, there were markedly different isolation rates of campylobacters between supermarket and retail market (P < 0.05). Enteropathogenic campylobacters (C. jejuni and C. coli) were found on 68% of whole chickens, 100% of chicken parts, and 100% of organs from retail markets. In supermarkets, the isolation rates of these campylobacters from whole chickens, chicken parts, and organs were 42%, 53%, and 60%, respectively. The low isolation rates of the two campylobacters isolated from chicken samples in supermarkets differed statistically from those obtained from traditional retail markets (P < 0.10). The API CAMPY test kit also was evaluated for the identification of Campylobacter spp. as compared with the conventional identification method. The results showed that the API CAMPY test kit (Biomerieux, Marcy-l'Etoile, France) could efficiently detect 87 Campylobacter spp. isolates from chicken samples examined, with 100% agreement at the genus level to 94% at the species level as compared with conventional methods.

Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other countries with long coastlines. This paper reports on the development of a new random amplified polymorphic DNA (RAPD) method for the molecular typing of this pathogen. The 10-mer primer 284 (5'-CAG GCG CAC A-3') was selected to generate polymorphic amplification profiles of the genomic DNA at an annealing temperature of 38 degrees C. A total of 308 clinical isolates of V. parahaemolyticus collected during food poisoning outbreaks in Taiwan, mostly occurring between 1993 and 1995, plus 11 environmental and clinical reference strains were analyzed by this RAPD method. A total of 41 polymorphic RAPD patterns were recognized, and these patterns were arbitrarily grouped into 16 types (A to P). Types A, B, C, D, and E were the major types, and subtypes C3, C5, E1, B1, D2, and A2 were the major patterns. The major types were phylogenetically more closely related to each other than to any of the minor types.

Vibrio parahaemolyticus is a prevalent food-borne pathogen in Taiwan, Japan and other Asian countries. This work presents a novel ribotyping method for the molecular epidemiological examination of this pathogen. Genomic DNA was fragmented by HindIII digestion and hybridized with cDNA probe for Escherichia coli 16S and 23S RNA genes. A total of 121 isolates obtained from outbreaks during 1992 and 1994 in Taiwan were characterized by this ribotyping method. Four to seventeen restricted fragments were visualized in these isolates. After hierarchical cluster analysis, these isolates were grouped into thirty different ribotypes. In addition, A3, A7, E3 and F1 were the major ribotypes, consisting of 22.3, 13.2, 9.1, and 8.3% of the isolates, respectively. A, E, F, G and B were the major groups, consisting of 46.2, 14.0, 9.1, 6.7, and 6.7% of the isolates, respectively. The discriminatory ability of this ribotyping method, as determined by Simpson's index of diversity, was 0.93, which closely resembled that of a previously reported pulsed-field gel electrophoresis method.

Vibrio parahaemolyticus is one of the most important food-borne pathogens in Taiwan, Japan, and other costal regions. We report on the development of a pulsed-field gel electrophoresis (PFGE) method for the molecular typing of this pathogen. Genomic DNA was digested with SfiI, and the fragments were resolved on 1% agarose with a contour-clamped homogeneous electric field apparatus set at 190 V and a pulse time of 3 to 80 s. A total of 130 selected isolates obtained from outbreaks during 1993 and 1994 on Taiwan were also characterized by this PFGE method. These isolates were grouped into 14 PFGE types which consisted on one to six patterns, and a total of 39 patterns were identified. Most of these domestic clinical isolates could be clustered into several major types (types A, B, C, and G). These major types showed relatively low degrees of similarity to several foreign strains and other domestic but environmental strains. Strain CCRC12863, which originated from Japan, was close to the group consisting of F, G, and H PFGE types, suggesting a clonal relationship between this Japanese strain and other domestic isolates.