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The extent of arsenic contamination in drinking water and its potential threat to human health have resulted in considerable research interest in the microbial species responsible for arsenic reduction. The arsenate reductase gene (arrA ), an important component of the microbial arsenate reduction system, has been widely used as a biomarker to study arsenate-reducing microorganisms. A new primer pair was designed and evaluated for quantitative PCR (qPCR) and high-throughput sequencing of thearrA gene, because currently available PCR primers are not suitable for these applications. The primers were evaluatedin silico and empirically tested for amplification ofarrA genes in clones and for amplification and high-throughput sequencing ofarrA genes from soil and groundwater samples.In silico , this primer pair matched (≥90% DNA identity) 86% ofarrA gene sequences from GenBank. Empirical evaluation showed successful amplification ofarrA gene clones of diverse phylogenetic groups, as well as amplification and high-throughput sequencing of independent soil and groundwater samples without preenrichment, suggesting that these primers are highly specific and can amplify a broad diversity ofarrA genes. ThearrA gene diversity from soil and groundwater samples from the Cache Valley Basin (CVB) in Utah was greater than anticipated. We observed a significant correlation betweenarrA gene abundance, quantified through qPCR, and reduced arsenic (AsIII ) concentrations in the groundwater samples. Furthermore, we demonstrated that these primers can be useful for studying the diversity of arsenate-reducing microbial communities and the ways in which their relative abundance in groundwater may be associated with different groundwater quality parameters.IMPORTANCE Arsenic is a major drinking water contaminant that threatens the health of millions of people worldwide. The extent of arsenic contamination and its potential threat to human health have resulted in considerable interest in the study of microbial species responsible for the reduction of arsenic, i.e., the conversion of AsV to AsIII . In this study, we developed a new primer pair to evaluate the diversity and abundance of arsenate-reducing microorganisms in soil and groundwater samples from the CVB in Utah. We observed significantarrA gene diversity in the CVB soil and groundwater samples, andarrA gene abundance was significantly correlated with the reduced arsenic (AsIII ) concentrations in the groundwater samples. We think that these primers are useful for studying the ecology of arsenate-reducing microorganisms in different environments.

New Arsenate Reductase Gene ( arrA ) PCR Primers for Diversity Assessment and Quantification in Environmental Samples