Triplicate Sequencing:
The European Consensus Group on clinical management of HIV-1 tropism testing recommends to undertake triplicate PCR amplification and sequencing. In geno2pheno[coreceptor] this can be done by using the batch-upload feature. Just store your triplicates in a fasta file and upload it on the input page. The system will generate a prediction for each of these sequences and report them in a table that can be downloaded in csv-format (which can be opened with Excel, for instance). The lowest FPR-score reported should then be used for interpretation. A value below 10% is predictive for an X4 virus whereas a value above 10% reflects an R5 virus. In case that only one sequence can be generated, the cutoff should be increased from 10% to 20%.
Recommendations by the European Consensus Group on clinical management of HIV-1 tropism testing
The recommendations by the European Consensus Group on clinical management of HIV-1 tropism testing can be found in the original article: European guidelines on the clinical management of HIV-1 tropism testing
Deutsch-Oesterreichische Leitlinien zur Tropismus-Testung
Die Deutsch-Oesterreichischen Leitlinien zur Tropismus-Testung koennen hier gefunden werden.