Global surveillance of influenza is critical for improvements in disease management and is especially important for early detection, rapid intervention, and a possible reduction of the impact of an influenza pandemic. Enhanced surveillance requires rapid, robust, and inexpensive analytical techniques capable of providing a detailed analysis of influenza virus strains. Low-density oligonucleotide microarrays with highly multiplexed "signatures" for influenza viruses offer many of the desired characteristics. However, the high mutability of the influenza virus represents a design challenge. In order for an influenza virus microarray to be of utility, it must provide information for a wide range of viral strains and lineages. The design and characterization of an influenza microarray, the FluChip-55 microarray, for the relatively rapid identification of influenza A virus subtypes H1N1, H3N2, and H5N1 are described here. In this work, a small set of sequences was carefully selected to exhibit broad coverage for the influenza A and B viruses currently circulating in the human population as well as the avian A/H5N1 virus that has become enzootic in poultry in Southeast Asia and that has recently spread to Europe. A complete assay involving extraction and amplification of the viral RNA was developed and tested. In a blind study of 72 influenza virus isolates, RNA from a wide range of influenza A and B viruses was amplified, hybridized, labeled with a fluorophore, and imaged. The entire analysis time was less than 12 h. The combined results for two assays provided the absolutely correct types and subtypes for an average of 72% of the isolates, the correct type and partially correct subtype information for 13% of the isolates, the correct type only for 10% of the isolates, false-negative signals for 4% of the isolates, and false-positive signals for 1% of the isolates. In the overwhelming majority of cases in which incomplete subtyping was observed, the failure was due to the nucleic acid amplification step rather than limitations in the microarray.

The design and characterization of a low-density microarray for subtyping influenza A is presented. The microarray consisted of 15 distinct oligonucleotides designed to target only the matrix gene segment of influenza A. An artificial neural network was utilized to automate microarray image interpretation. The neural network was trained to recognize fluorescence image patterns for 68 known influenza viruses and subsequently used to identify 53 unknowns in a blind study that included 39 human patient samples and 14 negative control samples. The assay exhibited a clinical sensitivity of 95% and clinical specificity of 92%.

Mixtures of CH(3)CN and H(2)O are the predominant solvent systems used in reversed-phase liquid chromatographic (RPLC) separations, as well as in a multitude of other applications. In addition, acetonitrile is the simplest model for an amphiphilic molecule possessing both organic and polar functional groups. Although many studies have focused on this solvent system, the general nature of the intermolecular interactions are not fully understood, and a microscopic description of the proposed microheterogeneity that exists is still not clearly established. In the present study, we measure the spin-lattice relaxation times (T(1)) of (14)N to determine reorientational correlation times (tau(c)) of CH(3)CN-H(2)O solvent mixtures over the entire binary composition range and at temperatures ranging from 25.0 to 80.0 degrees C. At all compositions, the microscopic observable, tau(c), is found to be directly proportional to the macroscopic solution viscosity when scaled for temperature (eta/T). This indicates that for a constant composition, this system's dynamics are well described by hydrodynamic theory on a microscopic level. These results suggest that under appropriate conditions, the measurement of changes in quadrupolar relaxation times is a reliable means of determining changes in solution viscosity. We stress the importance of this approach in systems not amenable to traditional viscosity measurements, such as those having species in interfacial regions. This approach is used to examine the changes in the interfacial solution viscosity of CH(3)CN-H(2)O mixtures in contact with a commercially available C(18)-bonded stationary phase. The measurements indicate that CH(3)CN is motionally hindered at the stationary phase surface. The surface affected CH(3)CN has a larger dependence of tau(c) on temperature than the bulk CH(3)CN, indicating greater changes in the interfacial viscosity as a function of temperature. Additionally, the bulk relaxation data show direct correlations to existing models of proposed regions of structure for CH(3)CN-H(2)O mixtures. Using a microscopic hydrodynamic approach, we show that, quite unexpectedly, each of the experimentally determined parameters in the viscosity correlation plots change simultaneously, and we propose that these are indicative of changes in the distribution of species for this microheterogeneous liquid system. Although distinct regions for the onset of microheterogeneity have previously been proposed, within the framework of a microscopic hydrodynamic model and the recently proposed model of Reimers and Hall,(1) the present data support the existence of a microheterogeneous solvent structure that varies continuously over the full range of temperatures and compositions examined.