US 11,807,908 B2
Genetic markers used for identifying benign and malignant pulmonary micro-nodules and the application thereof
Changming Cheng, Shanghai (CN); Chao Yang, Shanghai (CN); Ruiqin Ma, Shanghai (CN); and Yin Zhou, Shanghai (CN)
Assigned to Shanghai Biomedical Laboratory Co., Ltd., Shanghai (CN)
Filed by Shanghai Biomedical Laboratory, Co., Ltd., Shanghai (CN)
Filed on Nov. 20, 2018, as Appl. No. 16/196,163.
Application 16/196,163 is a continuation in part of application No. PCT/CN2017/083019, filed on May 4, 2017.
Claims priority of application No. 201610353398.6 (CN), filed on May 25, 2016; and application No. 201810661113.4 (CN), filed on Jun. 25, 2018.
Prior Publication US 2019/0078167 A1, Mar. 14, 2019
Int. Cl. C12Q 1/6886 (2018.01); C12Q 1/6851 (2018.01); C12Q 1/6837 (2018.01); G16H 50/20 (2018.01); G16B 45/00 (2019.01)
CPC C12Q 1/6886 (2013.01) [C12Q 1/6837 (2013.01); C12Q 1/6851 (2013.01); G16B 45/00 (2019.02); G16H 50/20 (2018.01); C12Q 2600/106 (2013.01); C12Q 2600/112 (2013.01); C12Q 2600/118 (2013.01); C12Q 2600/158 (2013.01); C12Q 2600/16 (2013.01)] 9 Claims
 
1. A composition comprising first, second, third, fourth, fifth, and sixth combinations, wherein each combination comprises primers and a probe useful for detecting differential expression of genes in the peripheral blood of micronodular lung carcinoma patients and patients without the micronodular lung carcinoma, wherein the genes are (i) HSP90AA1 gene, (ii) UQCRQ gene, (iii) NDUFB2 gene, (iv) RPL24 gene, (v) CKLF gene, and (vi) GLRX gene; wherein
a) the first combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 13, wherein the first primer can hybridize to HSP90AA1 cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 14, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a first probe, wherein the first probe comprises the sequence of SEQ ID NO: 37 and is fluorescently tagged and can hybridize to the second polynucleotide;
b) the second combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 15, wherein the first primer can hybridize to UQCRQ cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 16, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a second probe, wherein the probe can hybridize to the second polynucleotide;
c) the third combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 23, wherein the first primer can hybridize to NDUFB2 cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 24, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a third probe, wherein the probe can hybridize to the second polynucleotide;
d) the fourth combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 25, wherein the first primer can hybridize to RPL24 cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 26, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a fourth probe, wherein the probe can hybridize to the second polynucleotide;
e) the fifth combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 27, wherein the first primer can hybridize to CKLF cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 28, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a fifth probe, wherein the probe can hybridize to the second polynucleotide; and
f) the sixth combination comprises (i) a first primer comprising the sequence of SEQ ID NO: 35, wherein the first primer can hybridize to GLRX cDNA and be extended to provide a first polynucleotide; (ii) a second primer comprising the sequence of SEQ ID NO: 36, wherein the second primer can hybridize to the first polynucleotide and be extended to produce a second polynucleotide and (iii) a sixth probe, wherein the probe can hybridize to the second polynucleotide.