Intratypic Differentiation on positive isolates

1.    Scope

·       This SOP explains the steps to be followed in ITD laboratory during rRTPCR procedure.

2.    Definition

·       rRTPCR- Real Time Reverse Transcriptase Polymerase Chain Reaction 

3.    Procedure

Sequence of events

• L20B positive isolates (L+R+, LL+R+, R+L+R+, RR+L+R+) are subjected to rRT PCR.

• If both the arms are positive then only LR positive tubes are sent for rRT PCR.
• If only L20 B arm is positive then LR positive tube is sent for rRT PCR.
• If only RD arm is positive then RLR or RRLR positive tubes are sent for rRT PCR.
• Positive isolate by rRT PCR are tested as per following protocol:

o   Wild –isolates are sent for sequencing to GSL within 24 hours

• The results of Positive isolate by rRT PCR- ITD may be:

o   Wild Type

o   Vaccine type

o   NPEV

o   Wild and Vaccine Mixture