Abstract
Waterborne diseases have emerged as global health problems and their rapid and sensitive detection in environmental water samples is of great importance. Bacterial identification and enumeration in water samples is significant as it helps to maintain safe drinking water for public consumption. Culture‐based methods are laborious, time‐consuming, and yield false‐positive results, whereas viable but nonculturable (VBNCs) microorganisms cannot be recovered. Hence, numerous methods have been developed for rapid detection and quantification of waterborne pathogenic bacteria in water. These rapid methods can be classified into nucleic acid‐based, immunology‐based, and biosensor‐based detection methods. This review summarizes the principle and current state of rapid methods for the monitoring and detection of waterborne bacterial pathogens. Rapid methods outlined are polymerase chain reaction (PCR), digital droplet PCR, real‐time PCR, multiplex PCR, DNA microarray, Next‐generation sequencing (pyrosequencing, Illumina technology and genomics), and fluorescence in situ hybridization that are categorized as nucleic acid‐based methods. Enzyme‐linked immunosorbent assay (ELISA) and immunofluorescence are classified into immunology‐based methods. Optical, electrochemical, and mass‐based biosensors are grouped into biosensor‐based methods. Overall, these methods are sensitive, specific, time‐effective, and important in prevention and diagnosis of waterborne bacterial diseases.
Highlights
Waterborne diseases have become a major public health issue globally, which affect more than half the population of the developing world
Water quality for potable and recreational purposes has been exclusively evaluated based on the culture-dependent enumeration and detection of fecal indicator bacteria (FIB), for example, total coliforms, Escherichia coli, or Enterococci, a strategy practiced for decades as a “gold standard” in the assessment of microbial safety of water (Figueras and Borrego 2010)
Such limitations include the biases generated in polymerase chain reaction (PCR) amplification due to secondary structure of the resulting amplicons, false diversity generated from sequencing errors or chimera formation, and the choice of primers used to target different small subunit rRNA hypervariable regions (Quail et al 2012; Kozich et al 2013; Schirmer et al 2015)
Summary
Waterborne diseases have become a major public health issue globally, which affect more than half the population of the developing world. Methods for DNA extraction from such environmental samples are required to be developed for rapid detection and enumeration of waterborne bacteria. Enzymatic reaction is stopped by 10 min incubation at 65°C and WGA-amplified product is directly used for qPCR This method enabled the detection of 1.8 CFU of E. coli in 100 mL of environmental water sample within a span of 5 h. PCR allows the detection of a single bacterial pathogen which is present in water by targeting specific DNA sequence (Maheux et al 2011). A sensitive CRENAME method coupled with real-time PCR was reported for specific detection of E. coli/Shigella in potable water using TaqMan probe (Maheux et al 2011). Microarrays are powerful genomic technology that are widely used to study gene expression under different cell growth conditions, detect specific mutations in DNA sequences, and characterize microorganisms in the environmental samples (Lee et al 2008). Methods other than FISH have been efficiently used since the last decade to monitor VBNC in water bodies such as immunological techniques, RT-P CR, and the commercial kit LIVE/DEAD® BacLight TM assay (Barer and Bogosian 2004; Rowan 2004; Pinto et al 2011)
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