Discrimination of Ad3 or Ad7 from Ads of other types in one-step multiplex PCR of 115 direct clinical specimens that produced amplicons of consensus primers

Multiplex PCR resultNo. of specimens that previously grew the following Ad typea:Total no. of specimens
Ad3 (n = 28)Ad7 (n = 35)Ads of other types (n = 52)
Amplification by Ad3-specific primers261b027
Amplification by Ad7-specific primers2c35b2d39
  • a Ad type determined by neutralization test.

  • b One specimen amplified 502-bp, 311-bp, and 171-bp bands. These 502-bp and 311-bp amplimers produced by multiplex PCR of clinical specimens were confirmed by sequencing analysis to be specific to Ad3 and Ad7, respectively.

  • c These two specimens amplified only the Ad7 band (311-bp size) and the Ad common bands on multiplex PCR, and the 311-bp-sized products were compatible with Ad7 bands via sequencing analysis. However, both of the specimens exhibited Ad3 bands upon optimized PCR with Ad3F/Ad3R primers.

  • d Two specimens that previously grew Ad2 (n = 1) and Ad5 (n = 1) produced 311-bp amplimers, as well as 171-bp amplimers. The sequencing of 311-bp amplimers of these specimens revealed that the 311-bp amplimers were specific to Ad7. Also, additional sequencing analysis of the 301-bp bands produced by hex1deg and hex2deg by Allard et al. (4) revealed Ad2- and Ad5-specific products, respectively.