CDC Develops Multiplex Hepatitis Assay
The U.S. Centers for Disease Control and Prevention (CDC) has developed a multiplex polymerase chain reaction (PCR) assay capable of simultaneously detecting all five known hepatitis viruses, according to an abstract presented at the annual meeting of the American Association for the Study of Liver Diseases (Nov. 7-11, Boston). While the five viral hepatitides are […]
The U.S. Centers for Disease Control and Prevention (CDC) has developed a multiplex polymerase chain reaction (PCR) assay capable of simultaneously detecting all five known hepatitis viruses, according to an abstract presented at the annual meeting of the American Association for the Study of Liver Diseases (Nov. 7-11, Boston). While the five viral hepatitides are clinically indistinguishable, various testing algorithms are employed to determine their etiology. Nucleic acid testing (NAT) remains the gold standard for diagnosis of active and viremic stages of infection, the CDC says. The CDC researchers designed standardized TaqMan (Life Technologies) assays for simultaneous detection of RNA for hepatitis A, C, D, and E viruses (HAV, HCV, HDV, and HEV) and the DNA for hepatitis B virus (HBV). After individual PCR assays for each virus were optimized to run under identical experimental conditions, the assays’ performances were evaluated on TaqMan Array Cards (TAC) to detect the five viral genomes simultaneously. “The hepatitis TAC assay has great potential for simplifying laboratory testing of viral hepatitides,” Maja Kodani, Ph.D., an associate service fellow in CDC’s Division of Viral Hepatitis, tells DTET. “It was not developed to replace current hepatitis testing algorithms; instead, it was developed to address a need for rapid testing in situations where traditional testing may be challenging and expensive.” Based on 329 clinical specimens, the TAC assay was able to identify all positive samples for HCV, HDV, and HEV, yielding 100 percent sensitivity for those strains. The TAC assay positively identified 43 of 46 HAV-NAT positive samples (sensitivity, 94 percent) and 36 of 39 HBV-NAT positive samples (sensitivity 92 percent). No false positives were detected for HBV, HCV, HDV, or HEV in NAT-negative samples (specificity 100 percent), while for HAV-NAT positive samples, the TAC assay’ specificity was 93 percent. While the group says they are working to further optimize and validate the assay before it is transferred to public health laboratories for routine use, they believe there are additional opportunities to expand the utility of the panel beyond global health and disease surveillance. Kodani says, for example, future work will add more pathogens such as HIV and other sexually transmitted diseases to make the panel useful for donor blood and organ testing, as well as for prenatal testing. Takeaway: A multiplex PCR-based assay can simultaneously detect all five known hepatitis viruses. With the addition of other sexually transmitted diseases, future iterations of the panel may expand its utility beyond public health disease surveillance and into the realm of donor blood and organ testing and prenatal testing.