Studying The HBV Viral Strains And Occult Hepatitis B Virus Infection Biomarkers Of People Aged 2-14 In Urumqi

Objective To investigate the relationship between S gene segment variable site and OBI after the pupils(7-14 years old) and preschool children(2-6 years old) in fecting occult hepatitis B virus infection and the relationship between four kinds of mi RNA expression pattern and OBI, discuss the feasibility of mi RNA for early diagnosis of OBI. Method Urumqi county,s four primary school selected by cluster sampling method, using ELISA method(ELISA) to detect HBs Ag and anti-HBs serological index. The 469 primary school students, two indicators of HBs Ag and anti-HBs were negative, the HBV-DNA extracted by kit of QIAGEN company, polymerase chain reaction(PCR) amplified HBV-DNA S gene fragment, 43 valid objective bands of PCR amplified products detected by HBV S gene region(544bp) sequence. Establishing of HBV sequence database.By MEGA 5 soft to compare the sequencing results with the NCBI sequence database, in order to obtain nucleotide variation regularity of HBV region of the S gene; evolutionary selection model based on S fragment, determination of the evolutionary tree of the reference sequence and INSDC included, to determine the viral genotypes, compare the genotype distribution difference, analysis the nucleotide variation sites, in 40 cases of primary school sequencing success, according to the 1:1 analysis( the Han nationality, minority) pairing, a total of 20 for the OBI study. After cluster sampling, random select 20 healthy students and 20 students of positive HBs Ag as control, and matched with OBI group in age,gender and ethnicity. A total of 60 serum samples were detected by QIAGEN kit to extract RNA, transcript c DNA, determinate the relative expression amount of four mi RNA by fluorescence quantitative PCR. The choice of 4 kinder gardens in the sa-me way, using ELISA to detect HBs Ag, HBs Ab, HBe Ag, HBe Ab and HBc Ab in preschool children park, the HBV-DNA product was extracted with Promega kit, 259 cases of HBs Ag negative serum, PCR amplification of HBV-DNA S gene fragment, on the discovery of the 21 copies of the target band sequencing. Nucleotide variation obtained with the above method, to determine the gene type. Result In the 469 pupils, blood samples were amplified with PCR were found 43 HBV-DNA positive samples. Among them, 40 sequenced successfully, 32 cases of B genotype samples were serotype adw2, 5 samples were serotype adrq+ of 6 patients with C genotype, 1cases were serotype ayw2. 2 cases of D genotype samples were serotype ayw2. D g-enotype appeared only in the Kazak students. S gene sequence translated into the corresponding amino acid respectively, I110 L, S113 T, K122 R, T126 I, F134 Y, T143 S, K159 G and K160 R mutations.Differences in the amino acid of S gene mutation in the MHR District of gender, et-hnic was no statistical significance. Four kinds of miRNA in healthy controls and infected with HBV and healthy control subjects and patients of OBI expression, let-7-c, mi R-122 and mi R-150 was statistical difference significance, but no significant statistical difference of mi R-23 b in the serum. In addition, patients with OBI, let-7c, mi-R-23 b, mi R-122 and mi R-150 in the difference between the Han nationality and the minority nationalities have no statistical significance in serum. let-7c, mi R-122, m-i R-150 and mi R-23 b variants with the sequence of MHR gene region number is negative correlation,the correlation coefficients were-0.606,-0.902,-0.955 and-0.897, the P values were 0.202,0.009,0.003 and 0.015.Mi R-122, mi R-150, Let-7c and S were significantly related to amino acid mutation gene region MHR number.In the serum of three kinds of mi RNA expression differences to distinguish between OBI patients and normal controls, the area under the curve is 96.4%(sensitivity: 92.6%, specificity:90.8%). Receiver operating characteristic curve evaluate mi RNA, reflect certain diagnos-tic value. 259 samples of preschool children were found 21 HBV-DNA positive samples sequenced successfully. 1 cases due to the presence of amino acid insertion and leads to a frame shift mutation, so did not d-etect the gene type. The other 20 samples were C genotype, serotype were adrq+ sub-t ype. S gene sequence translated intoamino acids respectively I110 L, S123 T, T124 S,T126I, T131 N, F134 Y and P142 H mutations. 126 sites and 134 sites of amino acid variation in the pupils and preschool children was statistical difference significance(P<0.05). Conclusion In pupils and preschool children, after occult hepatitis B virus i-nfection, the S gene of amino acids have variation of different frequency. The expr-ession number of mi RNA and S region of the MHR gene of mutations of amino acid variation showed a certain correlation. Expression number of three mi RNA associated with OBI and reflected a certain diagnostic value.