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. Author manuscript; available in PMC: 2022 Nov 1.
Published in final edited form as: J Immunol Methods. 2021 Aug 28;498:113132. doi: 10.1016/j.jim.2021.113132

Table 4A.

Performance characteristics of RNA isolated from home processed filtrates vs. GEM Core processed filtrates.

Pt.no S.no Volume (cc)a A260/280 Ratiob Nanodrop based total RNA (μg)c Qubit based total RNA (μg)d 18S rRNA copies/ μg RNAg TGFb1 copies/ μg RNAg
Home Lab Home Lab Home Lab Home Lab Home Lab Home Lab
1 1 n/a 55 2.05 2.1 4.72 5.93 6.14 8.86 1.38E+09 1.07E+08 3.11E+03 1.68E+01
2 n/a 32.5 1.91 2.01 0.83 2.51 0.42 1.57 9.77E+08 1.42E+09 5.69E+02 2.23E+03
3 n/a 117.5 2.09 1.99 0.52 1.25 0.18 0.68 3.64E+08 5.49E+08 1.11E+02 9.36E+01
4 n/a 45 1.84 1.81 0.82 1.10 0.44 0.63 2.20E+09 4.32E+09 8.51E+03 9.51E+03
2 1 50 70 1.97 1.99 1.63 2.91 0.58 1.09 8.97E+08 5.54E+08 5.55E+02 8.65E+02
2 9.5 42.5 2.02 1.93 2.18 0.96 1.53 0.56 4.30E+08 3.56E+09 1.13E+02 7.58E+03
3 5 40 1.85 2.38 3.04 0.27 0.29 0.13 2.14E+09 4.32E+08 6.25E+00 1.70E+02
4 13 40 1.97 1.69 1.19 0.24 0.58 0.11 1.32E+09 3.91E+08 2.31E+02 4.10E+02
5 7 45 1.5 2.01 0.26 0.49 0.05 0.29 1.60E+08 5.29E+08 6.25E+00 2.82E+02
3 1 90 60 1.99 1.93 9.42 18.54 5.31 6.56 9.43E+08 1.06E+09 1.25E+03 1.00E+03
2 80 45 1.96 1.96 12.68 7.39 6.42 3.36 3.72E+09 1.61E+09 4.08E+03 1.29E+03
3 80 50 2.00 1.98 1.69 15.50 0.9 8.42 1.98E+08 1.77E+09 4.14E+01 1.45E+03
4 1 55 47.5 2.04 1.99 8.49 3.86 8.24 4.06 1.74E+08 2.56E+08 5.23E+02 6.61E+02
2 70 60 1.91 1.86 3.17 1.62 1.96 1.02 1.26E+08 7.26E+07 2.17E+02 4.80E+01
3 50 120 1.89 1.99 2.15 5.29 1.9 3.81 1.09E+08 2.06E+08 3.97E+02 1.15E+02
4 50 50 1.95 1.95 1.26 1.98 2.91 1.94 4.93E+08 8.03E+07 1.55E+03 1.04E+02
5 60 60 1.98 1.95 2.82 2.63 2.68 1.67 5.05E+08 1.83E+08 1.59E+03 2.67E+02
5e 1 n/a 75 2.03 2.01 3.16 1.73 2.50 1.29 1.88E+08 1.26E+08 2.80E+03 7.19E+02
2 50 45 2.01 2.03 1.16 2.26 0.87 1.75 1.28E+08 1.42E+08 9.42E+02 6.27E+02
3 60 52.5 1.76 2.07 0.55 4.97 0.55 5.87 1.61E+08 6.26E+07 8.36E+02 1.21E+02
4f 60 60 1.58 1.43 0.58 0.12 0.46 n/a 3.19E+08 2.76E+07 1.30E+03 1.07E+01
5 60 32.5 2.06 2.04 0.47 0.44 0.51 0.27 3.01E+08 1.03E+08 1.94E+03 1.54E+02
Median 55 50 1.97 1.99 1.66 2.12 0.89 1.57 3.97E+08 3.24E+08 7.03E+02 3.46E+02
25th Percentile 31.5 44.38 1.88 1.93 0.76 0.84 0.46 0.60 1.71E+08 1.06E+08 1.91E+02 1.12E+02
75th Percentile 65 60 2.02 2.02 3.16 5.05 2.74 3.94 1.06E+09 1.15E+09 1.68E+03 1.07E+03
Mann Whitney P value h 0.72 0.57 0.75 0.47 0.44 0.37

Kidney transplant recipients were trained by GEM Core Staff to perform initial processing of urine at home. The filtrates prepared by the patients (Home) were stored overnight at ambient temperature and brought to the transplant clinic the following day. The patients provided a freshly voided urine for the GEM Core staff to be processed at the Core. RNA from the filtrates prepared at home and the filtrates processed at the Core were stored at −80°C prior to total RNA isolation by the GEM Core Staff (Lab).

a

The volume of urine sample processed. Home volume was self-recorded by the kidney allograft recipients for 17 of the 22 urine samples and unavailable (n/a) for 5 home processed samples

b

A260/280 ratio measured using the NanoDrop® ND-1000 spectrophotometer.

c

The amount of RNA measured using the NanoDrop® ND-1000 spectrophotometer.

d

The amount of total RNA measured using Qubit fluorometer.

e

In one patient (Pt. no 5, shaded), reverse transcription of RNA to cDNA was based on NanoDrop® measurement and in the remaining 4 patients, reverse transcription of RNA to cDNA was based on Qubit measurement.

f

One urine sample (sample 4) from patient 5 did not have sufficient RNA (n/a) for measurement of RNA yield using Qubit Fluorometer.

g

Absolute copy number of 18S rRNA and TGF-β1 mRNA were measured with customized RT-PCR assays using gene specific oligonucleotide pairs and TaqMan probes on the 7500 Fast Real-Time PCR System.

h

P values derived using Mann-Whitney test.