Multiplex polymerase chain reaction-based reverse line blot hybridization to detect pathogens causing neonatal bacterial meningitis and relevant drug resistance genes / 中华围产医学杂志

Objective@#To evaluate the performance of multiplex polymerase chain reaction-based reverse line blot hybridization (mPCR/RLB) in the detection of pathogens causing neonatal bacterial meningitis and associated drug resistance genes.@*Methods@#Clinical data and cerebrospinal fluid (CSF) samples were collected retrospectively from 80 cases diagnosed with neonatal bacterial meningitis in Beijing Children's Hospital from January 1, 2012 to December 31, 2018. A total of 100 CSF samples were obtained including 80 samples collected after admission (12 before and 68 after antibiotic treatment) and 20 recollected at follow-up. All CSF samples were analyzed by conventional culture, susceptibility test and mPCR/RLB. Differences in the detection of pathogens and drug resistance genes were analyzed by Chi-square test.@*Results@#(1) Among the 80 first-collected CSF samples, mPCR/RLB revealed significantly higher positive rate than conventional culture [26.3% (21/80) vs 7.5% (6/80), χ2=10.025, P=0.002]. No significant difference was showed between the two methods in analyzing the 12 samples collected before antibiotic therapy (9/12 vs 5/12, χ2=1.543, P=0.214), while the positive rate in 68 samples collected after antibiotic intervention detected by mPCR/RLB was obviously higher than that by conventional culture [17.6% (12/68) vs 1.5% (1/68), χ2=13.176, P<0.001]. (2) Conventional culture results of the 20 samples collected during follow-up were all negative, but four were positive using mPCR/RLB, which were also positive previously. Furthermore, the results of both methods in previous detections were identical. (3) According to the conventional culture results, the pathogens were Escherichia coli (three cases), Group B Streptococcus (two cases) and Listeria monocytogenes (one case), while mPCR/RLB detected Escherichia coli (four cases), Group B Streptococcus (five cases), Listeria monocytogenes (four cases), Neisseria meningitidis (four cases), Haemophilus influenzae b (one case), Gram-negative bacteria (one case), Gram-positive bacteria (one case), and Listeria monocytogenes and Haemophilus influenzae b coinfection (one case) in 80 first-collected CSF samples. (4) Antibiotic susceptibility test showed that one Escherichia coli strain produced extended spectrum beta-lactamases. Drug resistance gene detection by mPCR/RLB showed that acrA, acrB, CTX-M (consistent with antibiotic susseptibility test) and TetM genes were positive in three, two, one and one case, respectively.@*Conclusions@#mPCR/RLB is of great clinical value due to its higher detection rate and better accuracy compared with bacterial culture and can also detect drug resistance genes.

Fetal Lateral Ventricular Dilatation Complicated with Malformation Diagnosed by Ultrasound and Its Relationship with Chromosome Abnormalities / 中国医学影像学杂志

Purpose To investigate the relationship between fetal lateral ventricle dilatation complicated with malformation and chromosome abnormalities,so as to provide reference for clinical consultation.Materials and Methods The ultrasound images,karyotype analysis and high resolution microarray comparative genomic hybridization (aCGH) results of 150 fetuses with lateral ventricular dilatation diagnosed by ultrasound were analyzed retrospectively.Results Among 150 lateral ventricular dilatation fetuses,81 cases were isolated lateral ventricle dilatation,30 cases were found complicated with fetal ultrasound soft index,22 cases with other CNS malformations and 17 cases with other malformations.Karyotype analysis of the above 4 groups showed 13 cases of abnormal karyotypes and 15 cases ofaCGH abnormalities.There was statistical significant difference (P＜0.05) in each group of abnormal chromosome and aCGH detection rate,in which the softer marker's group had a significantly higher rate than the isolated ventriculomegaly's group in abnormal chromosome and aCGH (P＜0.05).There was no statistical significant difference in any other groups (P＞0.05).Conclusion Fetal systems should be carefully examined when prenatal ultrasound reveals lateral ventricular dilatation.The probability of abnormal chromosome increases significantly when complicated with fetal ultrasound soft index or other structural abnormalities.Chromosomal abnormalities need to be excluded and regular ultrasound follow-up is necessary in fetuses with isolated lateral ventricles.

The application of multicolor probe melting curve analysis for the prenatal diagnosis ofβthalassemia / 中华检验医学杂志

Objective To establish a method of multicolor melting curve analysis for the prenatal diagnosis ofβthalassemia.Methods Methodology establishment.A total of 95 cases, including 9 fetal villi samples(10-13 weeks)and 86 amniotic fluid samples(18-24 weeks)were collected by Center for Prenatal Diagnosis of Shenzhen Maternity and Child Healthcare Hospital between January 2014 and December 2014.A double-blind test was done to detect the mutations of beta globin gene by means of reverse dot ( RDB) blot and multicolor melting curve analysis ( MMCA).The consistency of the two methods is compared.Results The results of 93 cases detected by MMCA and RDB are completely consistent.The results of the 2 cases detected by MMCA after correction are the same as the results detected by RDB.Finally, the coincidence rate of the result was 100%.Conclusion MMCA can be applied to the prenatal diagnosis ofβthalassemia as an effective supplement to RDB.

Biological warfare agent detection technology and research progress / 军事医学

As a weapon of mass destruction ,the biological weapon , composed of biological warfare agents and their re-lease devices,is characterized by strong pathogenicity , large pollution areas, various routes of infection, low cost, user-friendliness and a large number of impact factors .Although the United Nations has banned the use of biological weapons , there are still some countries and regions that continue biological weapon researches .In addition, illegal use of biological warfare agents in the field of terrorism and non-military arena poses a serious threat to public safety .Early detection of bio-logical warfare agent use and determination of its type are crucial to biological weapon defense and epidemic control .There-fore, to enhance researches on rapid detection and early warning of biological warfare agents is of great significance .This paper reviews the main technologies currently applied to the field of biological warfare agent detection and their progress .

Clinical investigation to compare aCGH and FISH in preimplantationgenetic diagnosis of chromosome translocation carriers / 中华妇产科杂志

Objective To investigate the clinical use of array comparative genomic hybridization (aCGH) with fluorescence in situ hybridization (FISH) in preimplantion genetic diagnosis (PGD)for reciprocal and Robertsonian translocation carriers.Methods From Jan.2012 to Jun.2013,a total of 220 PGD cycles from 151 reciprocal translocation and 62 Robertsonian translocation carrier couples,including 33 cycles for reciprocal translocation carriers and 22 cycles for Robertsonian translocation carriers performed using array CGH,and 119 cycles for reciprocal translocation carriers and 46 cycles for Robertsonian translocation carriers performed using FISH were retrospectively studied.The rate of accurate diagnosis was compared between two methods.Results Normal and/or balance rates of the two translocated chromosomes detected by aCGH for both reciprocal and Robertsonian translocation carriers were 38.20％ (123/322) and 67.20％ (127/189),significantly higher than 15.39％ (195/1 267) and 30.75％ (202/657) by FISH (all P ＜0.05).Abnormal rates of the two translocated chromosomes detected by aCGH for both reciprocal and Robertsonian translocation carriers were 59.32％ (191/322) and 30.69％ (58/189),significantly lower than 83.03％ (1 052/1 267) and 67.43％ (443/657) by FISH (all P ＜ 0.05).And the rate of aneu ploidy in non-translocated chromosome from reciprocal translocation embryos was 20.19％ (65/322),which was significantly lower than 38.62％ (13/189) from Robertsonian translocation embryos (P ＜ 0.01).Conclusions Normal and/or balance rates of the two translocated chromosomes detected by array CGH were significantly higher than FISH.And the rate of aneuploidy in non-translocated chromosomes from reciprocal translocation embryos was significantly lower than that from Robertsonian translocation embryos.

Clinical value of human papillomavirus genotyping testing in the diagnosis of cervical intraepithelial neoplasia / 中华妇产科杂志

Objective To Investigate the value of HPV genotyping in diagnosis of cervical intraepithelial neoplasia(CIN).Methods From July 2012 to February 2013,200 women from Tianjin Medical University General Hospital and 244 women from Affiliated Hospital of the Chinese People's Armed Forces Logistics College were selected to be HPV genotyping test and thin liqid-based cytology test.Consequently,132 samples were performed colposcopy test and cervical biopsy.Results HPV prevalence was 26.4％ (117/444) in this study.The infection of one type HPV was more common.The top 5 of HPV types were HPV16,58,33,18,and 52.The top 5 of the risk for CIN Ⅱ and above followed HPV16,33,39,52 and 18.There was no significant difference between age and HPV positive rate (x2 =0.948,P ＞ 0.05).Multiple infection and cervical lesions rank correlation analysis(r =0.132,P ＞0.05).For CIN Ⅱ and above disease,cytology positive rate was 90％ (44/49),and HPV positive rate was 96％ (47/49) cytology combine HPV positive rate was 98％ (48/49,x2 =0.063,P ＞ 0.05).Conclusions HPV infection should increasing trends with age.Cytology test and HPV genotyping test had good consistency.The combination of them can improve the sensitivity for high-grade lesions.

Appropriate technologies for molecular diagnostic in personalized medicine / 中华检验医学杂志

With the development of genomics , bioinformation , engineering , computer science and other fields, it has witnessed explosive growth of molecular diagnostic technologies.More and more technologies are used in desease diagnosis , therapy and prevention , which have presented a huge opportunity and challenge for clinical laboratory.Each technology has corresponding field of application , so it is a crucial problem for clinical laboratory to select appropriate molecular diagnostic technology for personalized medicine.

Comparison between probe melting curve analysis based on real-time fluorescent PCR and reverse dot blot assay for gene and prenatal diagnosis of β-thalassemia / 中华检验医学杂志

ObjectiveTo evaluate the reliability of the probe melting curve analysis (PMCA) based on real-time fluorescent PCR assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.MethodsA total of 135 cases of peripheral blood samples were collected from Zhuhai Municipal Maternity and Child Healthcare Hospital between 2008 and 2010.All the samples were performed preliminary diagnosis according to the hematological data.Of these,119 cases were diagnosed as β-thalassemia trait,4 cases were diagnosed as severe thalassemia and 12 cases were normal.In addition,44 cases of amniotic fluid and 8 cases of cord blood with high-risk for severe β-thalassemia were also collected.The diagnostic reliability of the PMCA assay and reverse dot blot assay was evaluated on 187 above-mentioned cases by direct DNA sequencing analysis in a double-blind study.ResultsThe genotypes of 185 cases were detected accurately based on the PMCA assay except for two cases:one heterozygote with Ini( ATG ＞ AGG) was omitted and another heterozygote couldn't be distinguished between CD43 ( G ＞ T) and CD37 ( G ＞ A ).For the RDB assay,only one heterozygote with CD71-72 ( + T) was not detected accurately in the above-mentioned cases.Compared with the DNA sequencing analysis,the sensitivity,specificity,negative predictive value,positive predictive value and diagnostic efficiency of the PMCA assay were 98.75％,100.00％,93.10％,100.00％ and 98.93％,respectively.The corresponding value of the RDB assay were 99.38％,100.00％,96.42％,100.00％ and 99.47％,respectively.There were no significant between-group differences in the diagnostic efficiency of the two assays ( P ＞ 0.05 ).The results of prenatal diagnosis were in complete concordance with the follow up results,after the birth or induced labour of the fetuses.ConclusionsThe PMCA assay can be used as an alternative and verified method of RDB assay for the genetic diagnosis and prenatal diagnosis of β-thalassemia.

The establishment of a gene microarray for detecting the common pathogenic fungi / 中华检验医学杂志

Objective To develop a gene microarray system for detection of clinical common pathogenic fungi.MethodsThere were 8 clinical common fungi chosen as the subjects including Candida albicans,Candida glabrata,Candida tropicalis,Candida parapsilokis,Candida pseudotropicalis,Aspergillus terreus,Aspergillus flavus,Aspergillus oryzae,Aspergillus fumigatus.Universal primers,probes and specific probes for the PCR amplification and microarray preparation were designed in ITS region of the fungi genomic DNA.The PCR products amplified from those fungi's genome DNA were denatured and hybridized with the probes in gene microarray.The rapid detection of fungi was based on the investigation on the fluorescent signal intensity in the chip.The detection results of gene microarray system were verified by true positive and negative clinical samples.Results There were totally 25 positive samples identified by clinical routine microbiological methods.The 10 samples identified as bacteria positive were determined as negative without fluorescent signal by the fungi gene microarray,while the 12 samples identified as fungi positive were determined as positive with certain fungus by the fungi gene microarray.And 3 artificial Candida krusei samples were detected as fungi positive,while they were failure to be identified as certain fungus.There was no fluorescent signal in positions of the 8 fungi specific probes,but there was fluorescent signal in the position of fungi universal probe.It indicated that there were fungi in the samples but it couldn't identify the species of the fungi,because the Candida krusei wasn't included in the detection fungi list of the fungi gene microarray.ConclusionsThe fungi gene microarray established by the study could detect the common fungi in clinic rapidly and accurately.This study lays technology foundation for clinical application of gene chip.

Influence of different amplification methods and probes with various lengths on the results of comparative genomic hybridization analysis of preimplanted single blastomere / 中华围产医学杂志

Objective To investigate the effect of different amplification methods and probes with various length on the results of comparative genomic hybridization (CGH) analysis of pre-implanted single blastomere and to establish the basis for preimplantation genetic diagnosis.Methods Twenty blastomeres of embryo at 6-8 cells stage were randomly divided into A and B group with 10 in each.Twenty peripheral blood lymphocytes from a healthy man were similarly divided into C and D group with 10 in each.Degenerate oligonucleotide primed polymerase chain reaction (DOP-PCR) was used to amplify whole genomic DNA in group A and C,and multiple displacement amplification (MDA) was used in group B and D for whole genome amplification (WGA).The specificity of resultant products was confirmed by amplification of TBX1 gene exon 2.CGH was performed respectively with 250-750 bp and 750-2000 bp probes prepared from the amplified whole genomic DNA.The result of CGH was verified by sex-determining region of Y (SRY).Results (1) Nine of the 10 samples in group A and all in group C were amplifiable by DOP-PCR,but there were multiple non-specific bands in the amplification of TBX1 exon 2 when WGA products were used as templates.When 250～750 bp probe was used in CGH,1 of the 5 blastomeres was failed and another one had different karyotype from that analyzed by SRY.(2) All samples in group B and D were successfully amplified by MDA,and the non-specific bands were significantly less in the amplification of TBX1 exon 2.All 5 blastomeres were successful in CGH with the 250～750 bp probe.Moreover,the karyotype was in agreement with that of SRY.(3) When 750 ～ 2000 bp probe was used,the CGH results were suboptimal.Conclusions In WGA of single blastomere,MDA is superior to DOP-PCR in the stability and specificity.The karyotype image detected by CGH with the 250～750 bp probe is clear and homogenous.

The chromosomal aberration detected by comparative genomic hybridization in lung cancer / 中国医师杂志

Objective To understand the molecular aberration at whole genomic level,CGH (comparative genomic hybridization) was used to investigate genetic abnormality in lung cancer.Methods Comparative genomic hybridization was performed in 17 cases to detect the global genomic aberration in cancer tissue cells.Results All of 17 cases detected by CGH showed chromosomal aberrations.The average numbers of chromosomal gains and losses in each case were 7.0 and 4.8 in NSCLC and 8.4 and 9.6 in SCLC,respectively.The frequency of gains and losses on chromosome had no significant differences between NSCLC and SCLC.The frequencies of gains on chromosomal arms 3q24 -28 and 11q13(58.3％ and 58.3％ ) in NSCLC were significantly higher than that in SCLC(0％ and 0％ ) ( P ＜0.05 and ＜0.05,respectively).Conclusions The cytogenetic aberration generally existed in lung cancer cells.Several regions ( more than one) of chromosomal aberration were involved in the carcinogenesis of NSCLC and SCLC.The regions and frequencies of chromosomal aberration in NSCLC were somewhat different from that in SCLC,which might result in the different biological behavior of the two types of lung cancer.The chromosomal aberration might be served as a marker to differentiate the two types of lung cancer.

Development of a compact disk type microfluidic chip based on DNA hybridization for phenylketonuria screening / 中华检验医学杂志

Objective To develop a phenylketonuria (PKU) screening method based on a compact disk (CD) type microfluidic chip capable of generating reciprocating flow within the microchannels that facilitate rapid DNA hybridization. Methods This microfluidic device consists of a two-layer structure: a polydimethylsiloxane (PDMS) top layer containing 12 DNA hybridization microchannels, and a bottom glass layer with immobilized hydrogel conjugated DNA arrays. The DNA arrays included R243Q, V245V and the blank control probes. When the CD device was spun, the PCR products were driven into the hybridization channel by centrifugal force. When the rotation of the CD device was stopped, capillary force pulled the PCR products solution to flow back to the channel. After the on-chip hybridization, the hybridization signals were captured on a fluorescence microscope. The specificity, detection limitation and reproducibility of this device were evaluated. Thirty DNA samples from pregnant women with suspected PKU were detected by this device.Then the results were compared with DNA sequencing results. Results With the compact disk type microfluidic chip, the hybridization time could be reduced to 15 min, sample consume could be as low as 1. 5 μl and the detection limitation was 0. 7 ng/μl. With the chip based method, samples of PKU patients and healthy controls were detected and the results were consistent with DNA sequencing results. Five different batches of chips and five micro-channels of each chip were selected to test one PKU patients with V245V mutation. All the results were positive, indicating good reproducibility. Four cases of V245V mutation and 1 case of R243Q mutation were found in 30 suspected PKU carried pregnant women. Conclusion The compact disk microfluidic device has advantages of simple, rapid and highly sensitive, thus is well suited to PKU screening.

Application of array-based comparative genomic hybridization in primary amenorrhea women / 中华检验医学杂志

Objective To explore the molecular mechanisms of primary amenorrhea by using arrayCGH technology. Methods Ten patients with primary amenorrhea and 10 female volunteers with regular menstrual cycles as healthy controls were selected. All patients and control samples were analyzed by conventional chromosome analysis (G-banding technology) and array-CGH technology, respectively. ArrayCGH was performed using Affymetrix Cytogenetic 2. 7M arrays following the manufacturer's standard protocol. Results Both the patient group and control group analyzed by conventional G-banding karyotype technology showed a negative result with a normal female karyotype: 46, XX. The result of array-CGH analysis demonstrated a microdeletion of approximately 110 000 bp located at the end of the short arm of X chromosome [46, X, del (X) (p22. 33 )] were identified in 5 patients, which was not detected in the control group. All healthy control samples by array-CGH analysis showed no pathological DNA copy number variation. Conclusions Array-CGH technology can improve the diagnosis rate of chromosomal disease at the DNA level. It is necessary to provide array-CGH for higher resolution genetic analysis of idiopathic primary amenorrhea patient who can not be identified by conventional technology.

Prenatal diagnosis by array-based comparative genomic hybridization in the clinical laboratory setting / 北京大学学报(医学版)

Array-based comparative genomic hybridization (array CGH), a method used to detect gains or losses of genetic material, has recently been applied to prenatal diagnosis of genomic imbalance in the clinical laboratory setting. This new and exciting diagnostic tool represents a major technological step forward in cytogenetic testing and addresses many of the limitations of current cytogenetic methods.Conventional chromosome analysis, the current gold standard in prenatal diagnosis, focuses primarily on the detection of common aneuploidies and is limited by its capacity to detect only those copy number changes that are large enough to be microscopically visible (typically 5-6 Mb in size at the 500 band level). In contrast, array CGH analysis simultaneously evaluates regions across the entire genome and al-lows for detection of unbalanced structural and numerical chromosome abnormalities of less than one hun-dred kb. Array CGH analysis also overcomes some of the limitations of chromosome analysis, such as the requirement for cell culture and longer reporting time, by using direct uncultured fetal specimens. With many diagnostic laboratories now embracing this technology, the past year has seen tremendous growth in the use of array CGH analysis for prenatal diagnosis. This review aims to summarize array CGH methodology and its current applications in prenatal diagnosis.

The Chromosomal Aberrations in the Head and Neck Squamous Cell Carcinoma Cell Lines:PCI-1, PCI-13 and PCI-50 / 대한이비인후과학회지

BACKGROUND AND OBJECTIVES: The knowledge about chromosomal aberrations manifestated in cancer has been spotlighted recently. The genetic analyses based on the knowledge about chromosomal aberrations are important for the development of diagnotic methods and evaluation of prognostic factors in cancers. Comparative genomic hybridization (CGH) is a powerful tool for evaluating chromosomal aberrations and array CGH significantly enhances these diagnostic effectiveness. The incidence of head and neck squamous cell carcinoma (HNSCC) has been increasing worldwide but the treatment outcomes still have been limited. The aim of this study is to evaluate the location of chromosomal aberrations in HNSCC cell lines with the combination of CGH and array CGH. Materials and MethodZZThe locations of chromosomal aberrations were evaluated in 3 HNSCC cell lines (PCI-1, PCI-13, PCI-50) using the combination of CGH and array CGH. RESULTS: The sites of chromosomal gain shown by CGH in all 3 cell lines were 8q22-qter, 9q32- qter, 10q22, 10q26-qter, 16q12.1-qter, 17p10-p13, 17q21-qter, 19p13.2-pter and 20q. The chromsomal loss found in 2 cell lines were 3p, 4q21-qter, and 18q21-qter. In array-CGH, gained loci were AHRR, MYT1 and PTGIS, etc. Loci of genetic losses were ELAVL4 and GRM7. CONCLUSION: In this study, we identified various genetic gains and losses using CGH and high resolution array-CGH. These data about the patterns of chromosomal aberrations in HNSCC cell lines would be a basic step for understanding more detailed genetic events in the carcinogenesis. CGH combined with array CGH can be a powerful option for transitional oncologic research.

Relationship between human papillomavirns and cervical carcinoma / 中华全科医师杂志

Objective To explore the relationship between high-risk human papillomavirus (HPV) DNA and biological behavior of cervical carcinoma. Methods Sixty-six patients of cervical carcinoma with cytological examinations and 103 patients of cervical carcinoma followed-up after surgical operation were selected for high-risk HPV DNA test with second-generation hybrid capture technique (HC2 Ⅱ). Results ①HPV DNA was positive in 62 and negative in four of 66 patients of cervical carcinoma with an overall prevalence of 94%. ②There was no significant difference in positive HPV DNA of patients with cervical carcinoma between their varied clinical stages and pathologic grades. But, HPV positivity and HPV DNA load in patients with myometrial invasion were higher than those in patients without invasion (P < 0. 05).③ HPV DNA conversed to negative in 99 of 103 patients (96%) with cervical carcinoma after surgical operation from positivity before operation. Conclusions High-risk HPV infection may correlate with angiogenesis, invasion and metastasis of cervical carcinoma and HC2 Ⅱ can be used as an effective method to detect HPV DNA.

Genetic analysis of chorionic villi specimen in spontaneous abortion using various methods / 中华妇产科杂志

Objective To explore the efficacy of multiplex ligation-dependent probe amplification (MLPA)combined with fluorescence in situ hybridization(FISH)and comparative genomie hybridization (CGH)combined with FISH in genetic analysis of chorionic villi specimen(CVS)of spontaneous abortion.Methods CGH+FISH and MLPA+FISH were used for genetic analysis of 29 CVS from spontaneous abortion and 6 normal CVS from selective abortion,in the mean time,those results were compared with conventional eytogenetic karyotyping.Results The report time were 40 hours in MLPA+FISH and 120 houm in CGH+FISH.The mean time of chorionic villi culture was(240±72)hours.The successful rate of specimen analysis were 97%(34/35)in CGH,100%(35/35)in MLPA,100%(35/35)in FISH and 91%(32/35)in conventional cytogenetic karyotyping.Apart from 1 case failed in CGH analysis,the results from MLPA+FISH were almost similar to that from CGH+FISH,however,that 1 specimen failed in CGH were detected successfully by MLPA+FISH.The discrepancy rate were 13%(4/31)in CGH+FISH and 12%(4/32)in MLPA+FISH respectively when compared with conventional cytogenetic analysis.Conclusions MLPA+FISH analysis present shorter detecting time and achieve 100%tale of successful report.This combined method was an important adjuvant approach to conventional cytogenetic karyotyping in CVS from spontaneous abortion.

Application of flow-through hybrization and gene chipon human papiliomavious subtype detection in Chongqing / 中国基层医药

Objective To evaluate the application of flow-through rapid hybridization technique and gene chip (HybriMax) on human papiUomavions (HPV) subtype in Chongqing.Methods Cervical tissue samples were taken under the colpnscope form 473 females who had cervical lesion for pathological analysis.The predictive value of HybriMax in cervical abnormality was compared with pathological results,which were used as golden standard.Resuits 13 different subtypes were found and total HPV positive rate was 63.0% (284/473) Among the 17 different subtypes ,the higher positive rote HPV subtypes were HPV16 (23.7%,112/473),HPV58 (12.7% ,60/473),HPV53(7.4% ,35/473).The HPV infection rates were higher with the worse d cervical lesion(X2=77.06,P＜0.01).Conclusions The most frequent subtypes of HPV infection in Chongqing cervical lesion were HPV 16,58.HybriMax was an effective method to detect HPV subtype in clinical.

The exploratory development of the related genes with rifampin and isoniazid in Mycobacterium tuberculosis / 中华检验医学杂志

Objective To detect the related genes with rifampin and isoniazid in Mycobacterium tuberculosis in sputums using the DNA chip technique and evaluate the fensibility of the clinical application of the DNA chip technique.Methods 586 sputum smear specimen was detected using the L-J cultivation to determine their drug resistance.Simultaneously.DNA chip was employed to detect the mutation of the frequent mutable points rpoB,katG/inhA in mycobaeterium tuberculosis isolates.These two assays were compared and samples showing discrepancy were chosen for additional sequencing to evaluate the accuracy of the detection.Results(1)There were 584 culture positive sputum smear specimens including 3(+)163 specimens,2(+)204 specimens,and 1(+)217 specimens.The drug fast results displayed that 361 strains were sensitive to INH,223 strains tolerated INH in which 93 strains tolerated it in low concentration while sensitive to it in high concentration.and 130 strains tolerated it in both low and high concentration.While 327 strains were sensitive to RFP.247 strains tolerated RFP in which 59 strains tolemted it in low concentration while sensitive to it in high concentration,and 188 strains tolerated it in both low and high concentration.(2)There were 367 positive strains(62.8%)and 217 negative strains(37.2%)identified by PCR amplification of the specific resistance gene fragments.The detection rate of the katG/inhA was 28.4%,and the mutation sites were mainly focused on the katG315(89.8%).The detection rate of the rpoB was 55.9%(137/247),and the mutation sites were mainly focused on rpoB531(68.6%)and rpoB 526(16.1%).(3)The sequencing of sample,which showed discrepancy with L-J cultivation and the DNA chip confirm a certain omission ratio.Conclusions It is feasible to detect the related resistant genes in Mycobacterium tuberculosis isolates using the DNA chip technique.The key factor is to raise the efficiency of the DNA extraction,the effciency of the PCR and the quality control of the experiment to facilitate its clinical application.