The second-line drug resistance characterization of Mycobacterium tuberculosis in patients with acquired immune deficiency syndrome and tuberculosis

doi:10.3877/cma.j.issn.1674-1358.2016.02.008

Abstract

Abstract:

Objective

To explore the second-line drug resistance and molecular characterization of Mycobacterium tuberculosis in patients with acquired immune deficiency syndrome (AIDS) and tuberculosis (TB).

Methods

Specimens of patients with AIDS and TB, hospitalized in Beijing Ditan Hospital, Peking University Teaching Hospital from April 2010 to Octomber 2012, were collected and sent to CDC for identification. Four first-line drugs (isoniazid, rifampicin, streptomycin and ethambutol) and four second-line drugs (capreomycin, kanamycin, ofloxacin and ethionamide) susceptibility tests were taken and gyrA, gyrB, rrs, tlya, eis, ethA loci were detected for genetic polymorphisms.

Results

Total of 31 Mycobacterium tuberculosis strains were identified, while 12 isolates showed resistance to capreomycin, 8 strains resistance to ofloxacin, 4 strains resistance to kanamycin and 5 strains resistance to ethionamide. The drug resistance rates were 38.71%, 25.81%, 12.90% and 16.13%, respectively. Besides, 7 multidrug-resistant isolates and one extensively drug-resistant isolate were identified and the drug resistance rates were 22.58% and 3.23%, respectively. The most frequent mutation loci were rrs₁₄₀₁, gyrA₉₄ and gyrA₉₀. Ofloxacin drug resistance rate was significantly higher in isolates resistant to first-line drugs than first-line drugs sensitive isolates (P = 0.012). Gender was not associated with drug resistance (P = 0.533). Ofloxacin drug resistance rate was significantly lower in > 40 years old group than the other two groups (P = 0.043). There was no drug resistance and CD4 level difference between initial treated group and retreated group (P = 0.333, 0.307).

Conclusions

There existed primary drug resistance of second-line anti-tuberculosis drugs in patients with AIDS and TB. Capreomycin drug resistance rate was the highest followed by ofloxacin.

Key words: Acquired immune deficiency syndrome, Tuberculosis, Drug resistance, Molecular characterization, Second-line drug

Ling Zhang, Yue Sun, Yong Chen, Guiju Gao, Siyuan Yang, Kanglin Wan, Xingwang Li. The second-line drug resistance characterization of Mycobacterium tuberculosis in patients with acquired immune deficiency syndrome and tuberculosis[J]. Chinese Journal of Experimental and Clinical Infectious Diseases(Electronic Edition), 2016, 10(02): 166-172.

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References 35

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引物名称	引物序列（5'→3'）
16s rRNAF	ACGGTGGGTACTAGGTGTGGGTTTC
16s rRNAR	TCTGCGATTACTAGCGACTCCGACTTCA
IS1561’F	CGTGGGTGGGCCCTGGATACGTGAACTCT
IS1561’R	AACTGCTCACCCTGGCCACCACCATTGACT
RV1510F	GTGCGCTCCACCCAAATAGTTGC
RV1510R	TGTCGACCTGGGGCACAAATCAGTC
RV1970F	GCGCAGCTGCCGGATGTCAAC
RV1970R	CGCCGGCAGCCTCACGAAATG
RV3877/8F	CGACGGGTCTGACGGCCAAACTCATC
RV3877/8R	CTTGCTCGGTGGCCGGTTTTTCAGC
RV3120F	GTCGGCGATAGACCATGAGTCCGTCTCCAT
RV3120R	GCGAAAAGTGGGCGGATGCCAGAATAGT
RV0577F	ATGCCCAAGAGAAGCGAATACAGGCAA
RV0577R	CTATTGCTGCGGTGCGGGCTTCAA

引物名称	引物序列（5'→3'）
16s rRNAF	ACGGTGGGTACTAGGTGTGGGTTTC
16s rRNAR	TCTGCGATTACTAGCGACTCCGACTTCA
IS1561’F	CGTGGGTGGGCCCTGGATACGTGAACTCT
IS1561’R	AACTGCTCACCCTGGCCACCACCATTGACT
RV1510F	GTGCGCTCCACCCAAATAGTTGC
RV1510R	TGTCGACCTGGGGCACAAATCAGTC
RV1970F	GCGCAGCTGCCGGATGTCAAC
RV1970R	CGCCGGCAGCCTCACGAAATG
RV3877/8F	CGACGGGTCTGACGGCCAAACTCATC
RV3877/8R	CTTGCTCGGTGGCCGGTTTTTCAGC
RV3120F	GTCGGCGATAGACCATGAGTCCGTCTCCAT
RV3120R	GCGAAAAGTGGGCGGATGCCAGAATAGT
RV0577F	ATGCCCAAGAGAAGCGAATACAGGCAA
RV0577R	CTATTGCTGCGGTGCGGGCTTCAA

引物名称	引物序列（5'→3'）	退火温度（℃）	目的片段长度（bp）
rrs-F	GCCGCGAGGTTAAGCGAAT	66	352
rrs-R	CGCCCACTACAGACAAGAAC
tlya1-F	AGGCGCACGAGGTGTTGTTG	66	527
tlya1-R	AACGACAGGTCGGCCACTACCAGGT
tlya2-F	ATGTCGGATACGGCCAGCTG	68	555
tlya2-R	ACTTTTTCTACGCGCCGTGC
eis-F	GCGTAACGTCACGGCGAAATTC	59	567
eis-R	GTCAGCTCATGCAAGGTG
gyrA-F	ATGCAGCGCAGCTACATC	60	509
gyrA-R	TTGTGCGGCGGGATATTG
gyrB-F	CCGCTGTGATCTCGGTGAAG	64	775
gyrB-R	AGACCCTTGTACCGCTGAATG
ETH1-F	ACTACAACCCCTGGGACC	65	667
ETH1-R	ATCATCGTCGTCTGACTATGG
ETH2-F	CCTCGAGTACGTCAAGAGCAC	65	692
ETH2-R	CCTCGACCTTCCCGTGA
ETH3-F	CGTTGACGGCCTCGACATTAC	68	342
ETH3-R	GGTGGAACCGGATATGCCTG

引物名称	引物序列（5'→3'）	退火温度（℃）	目的片段长度（bp）
rrs-F	GCCGCGAGGTTAAGCGAAT	66	352
rrs-R	CGCCCACTACAGACAAGAAC
tlya1-F	AGGCGCACGAGGTGTTGTTG	66	527
tlya1-R	AACGACAGGTCGGCCACTACCAGGT
tlya2-F	ATGTCGGATACGGCCAGCTG	68	555
tlya2-R	ACTTTTTCTACGCGCCGTGC
eis-F	GCGTAACGTCACGGCGAAATTC	59	567
eis-R	GTCAGCTCATGCAAGGTG
gyrA-F	ATGCAGCGCAGCTACATC	60	509
gyrA-R	TTGTGCGGCGGGATATTG
gyrB-F	CCGCTGTGATCTCGGTGAAG	64	775
gyrB-R	AGACCCTTGTACCGCTGAATG
ETH1-F	ACTACAACCCCTGGGACC	65	667
ETH1-R	ATCATCGTCGTCTGACTATGG
ETH2-F	CCTCGAGTACGTCAAGAGCAC	65	692
ETH2-R	CCTCGACCTTCCCGTGA
ETH3-F	CGTTGACGGCCTCGACATTAC	68	342
ETH3-R	GGTGGAACCGGATATGCCTG

标本类型	阳性例数	百分比（%）
血培养阳性	13	41.94
淋巴结分泌物	11	35.48
痰培养	10	32.26
肺泡灌洗液培养	4	12.90
腹水	2	6.45
脑脊液	1	3.23

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