Na stm 1

The recombinant allergens of P. aztecus, (Pen a 1) were purchased from Indoor Biotechnologies (Product Code: NA-STM-1) (Indoor Biotechnologies, Bangalore, India).
The cDNA of Der p 10 was synthesized using primer dT₁₇ and MMLV reverse transcriptase (Invitrogen, Massachusetts, USA) from total mRNA of D. pteronyssinus. The specific primers of Der p 10 were designed based on the accession number, AF016278.1^{50 (link)}, provided in the National Center for Biotechnology Information. The polymerase chain reaction (PCR) products of Der p 10 with six histidine (His) were ligated into a pQE30 vector and transformed into Escherichia. coli- M15. The selection was performed using kanamycin (25 μg/ml) and ampicillin (100 μg/ml). The 6 × His tagged recombinant proteins were induced using 1 mM isopropyl-β-D- thiogalactopyranoside (Promega, Wisconsin, USA). The proteins were purified by nickel-nitrilotriacetic acid agarose metal affinity column chromatography under native conditions. The results of purified rDer p 10 are shown in the Supplementary Fig. 1.

Carolina shrimp tropomyosin standard, monoclonal, and polyclonal anti-shrimp tropomyosin antibodies were purchased from Indoor Biotechnologies (product codes: NA-STM-1, MA-1A6, and PA-SHM, respectively; Cardiff, UK). Unconjugated and alkaline phosphatase-conjugated goat anti-rabbit antibodies, alkaline phosphatase-conjugated rabbit anti-mouse antibodies, and low-molecular-weight salmon sperm DNA were purchased from Sigma-Aldrich (St. Louis, MO, USA). Bovine serum albumin was purchased from PAN-Biotech (Aidenbach, Germany). p-Nitrophenylphosphate disodium salt was purchased from Serva (Heidelberg, Germany). Synthetic oligonucleotide and forward and reverse primers were purchased from Integrated DNA Technologies (Coralville, IA, USA). PCR reaction cleanup kit and nuclease-free water were purchased from Qiagen (Hilden, Germany). Oligonucleotide antibody conjugation kit was purchased from Abcam (ab218260, Cambridge, UK). Polycarbonate PCR strips were purchased from Roboscreen (Leipzig, Germany). High protein-binding 96-well plates (NUNC Maxisorp), Taq DNA polymerase, and SYBR Green PCR master mix were purchased from Thermo Fisher Scientific (Waltham, MA, USA). All other chemicals were of analytical grade and purchased from Sigma-Aldrich (St. Louis, MO, USA).