# Simultaneous Detection of Nine Key Bacterial Respiratory Pathogens Using Luminex xTAG(r) Technology
## 2 Materials and Methods
### Primers for Pathogen Amplification
\begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline \multirow{2}{*}{**Organism**} & \multirow{2}{*}{\begin{tabular}{c} **Target** \\ **Gene** \\ **Sequences** \\ \end{tabular} } & \begin{tabular}{c} **No. of Beads** \\ **Coupled with** \\ **“Anti-TAG”** \\ **Sequences** \\ \end{tabular} & \multirow{2}{*}{\begin{tabular}{c} **Sequences** (\(\mathbf{5^{\prime}}\)-\(\mathbf{5^{\prime}}\)) \\ **(\(\mathbf{\mu}\)M)** \\ \end{tabular} } & \multirow{2}{*}{\begin{tabular}{c} **Working** \\ **Concentration** \\ **(\(\mathbf{\mu}\)M)** \\ \end{tabular} } & \multirow{2}{*}{\begin{tabular}{c} **Length** \\ **(\(\mathbf{\mu}\)M)** \\ \end{tabular} } & \multirow{2}{*}{\
\begin{tabular}{c} **Detection** \\ **Limits** \\ \end{tabular} } & \multirow{2}{*}{
\begin{tabular}{c} **Source** \\ **Coments** \\ \end{tabular} } \\  & & & & & & & \\ \hline \multirow{2}{*}{_S. pneumoniae_} & \multirow{2}{*}{_qpsA_} & \multirow{2}{*}{21} & F-TCAAACTCTCAATTTTCATAAAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{350} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[19]} \\  & & & ACCCAATGCAGCTGTGAC & & & & \\  & & & R-biotin-GATGCGCGCAGACCTGAAT & & & & \\ \hline \multirow{2}{*}{_M. catarrallis_} & \multirow{2}{*}{_165 rRNA_} & \multirow{2}{*}{8} & F-AAAAATACTACTATTTCACTAA-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{140} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[20]} \\  & & & TIGGCCTGTGTGAAAAT & & & & \\  & & & R-biotin-GATGCGCGCGTGT & & & & \\ \hline \multirow{2}{*}{_S. aureus_} & \multirow{2}{*}{_nuc_} & \multirow{2}{*}{12} & F-CAATATTCAAATTTCAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{270} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[21, 22]} \\  & & & GCGATTTGGTGTAGCGTT & & & & \\  & & & R-biotin-AGCCAAGCCTTGCAGAATAAGC & & & & \\ \hline \multirow{2}{*}{_S. pyogenes_} & \multirow{2}{*}{_dunseB_} & \multirow{2}{*}{9} & F-CACATATATATTATTCAAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{93} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[23]} \\  & & & TCATCTGAGAAGCTCTGT & & & & \\  & & & R-biotin-TCGGTGTAGCCATATGCGTGT & & & & \\ \hline \multirow{2}{*}{_H. influenzae_} & \multirow{2}{*}{_p6_} & \multirow{2}{*}{8} & F-AAAAATACTACTATTTCACTTA-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{296} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[24]} \\  & & & TIGCGCGCGGTGTGTGT & & & & \\  & & & R-biotin-TGCGCGCGTGTGT & & & \\ \hline \multirow{2}{*}{_M. pneumoniae_} & \multirow{2}{*}{_P1_} & \multirow{2}{*}{12} & F-CAATATATTCAAATTTCAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{209} & \multirow{2}{*}{1 pg} & \multirow{2}{*}{[25, 26]} \\  & & & GCCACACCTGGGGGCGCGCGCGCGC & & & & \\  & & & R-biotin-GATGCGCGCGCGCGCGCGCGCGCGCGC & & & & \\ \hline \multirow{2}{*}{_Legionella_ spp.} & \multirow{2}{*}{_165 rRNA_} & \multirow{2}{*}{9} & F-CACATATATATATATATATATAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{650} & \multirow{2}{*}{1 pg} & \multirow{2}{*}{[27]} \\  & & & AGAATATGCCTGCTGCTG & & & & \\  & & & R-biotin-GTCAACATATGCGCGTGTGT & & & \\ \hline \multirow{2}{*}{_P. aeruginosa_} & \multirow{2}{*}{_qpr1_} & \multirow{2}{*}{14} & F-AAATTTCTCTTTTTCATCAAT-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{249} & \multirow{2}{*}{1 pg} & \multirow{2}{*}{[28]} \\  & & & ATCACACAATCGTCGAATATTCGTGT & & & \\  & & & R-biotin-CTGCGCGCGCGTGTTTTCCAG & & & \\ \hline \multirow{2}{*}{_K. pneumoniae_} & \multirow{2}{*}{_Mdh_} & \multirow{2}{*}{15} & F-TACTCTTACTAAATTCAAC-12C- & \multirow{2}{*}{0.3} & \multirow{2}{*}{364} & \multirow{2}{*}{0.1 pg} & \multirow{2}{*}{[12, 29]} \\  & & & GCGCGCGCGAGATCTAAGCT & & & & \\  & & R-biotin-TTCAGTCGCGCCGCACAAAGCTA & & & & \\ \hline \hline \end{tabular}
\end{table} 
Table 2: List of target bacteria, genes, primers, and their amplicon sizes.