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The RNA of the biological replicates was pooled, lyophilized, and stored using the RNAstable® Tube Kit (Biomatrica, San Diego, CA, USA).
The total RNA samples obtained from T. reesei were lyophilized and stored using the RNAstable tube kit (Biomatrica, San Diego, CA, USA) in order to maintain the RNAs integrity for sequencing.
Specimens were collected using swabs and inserted into sample collection tube (kit of 3 ml UTM™ medium with 2 polyester swabs, Copan Diagnostics Inc., Murrieta, CA, USA) and were shipped the same day of collection at a temperature of 2°C to −4°C.
Approximately 1 ml of venous blood was aliquoted into each of three separate tubes of the QuantiFERON-TB Gold In Tube kit (QFT-GIT) (Cellestis, Australia): one tube containing only heparin, a negative control (Nil), another tube containing phytohaemagglutinin (PHA), a positive control (Mitogen) and the third tube containing overlapping peptides of ESAT-6, CFP-10 and TB7.7 (M. tuberculosis antigen).
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QuantiFERON-TB Gold In tube kits were provided in part by Inverness Medical Ibérica SAU, the distributor of Cellestis in Spain; and T.SPOT-TB was provided in part by Oxford Immunotec by means of a Material Transfer Agreement.
PCR was carried out with a 16S rRNA primer set specific for M. pneumoniae [28] using the EasyStart™ 100 PCR mix-in-a-tube kit (Molecular Bio-Products, San Diego, CA).
The glass capillaries of 0.5 mm in diameter and 60 mm in length from Crystal-Tube Kit (Confocal Science Inc).
Capillary C-Tube Crystal-Tube Kit was purchased from Confocal Science Inc. Geobacillus zalihae strain T1 lipase was overexpressed in pGEX vector in the prokaryotic system.
The site for placement of PEG- tube was located via trans-illumination on the abdominal wall, followed by an incision, and placement of a cannula (provided in the PEG-tube kit).
The level of IFN-γ was measured using the QFT-GIT ELISA In-Tube Kit (Cellestis, Australia) and test interpretation and results were analyzed using QuantiFERON-TB Gold Analysis Software (Version 2.50, Cellestis, Australia).
The blood spot analyses were done using the IMMUCHEM™ NEONATAL TSH-MW ELISA (bulk kit −20 plates) and RIA kits (500 tubes kit) provided by MP Biochemicals, USA.
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