Age-related Reference Intervals for Total Collagen-I-N-terminal Propeptide in Healthy Korean Population

BACKGROUND: Procollagen type I N-terminal propeptide (PINP) is one of the most clinically useful bone formation biomarkers. Therefore, the purpose of this study was to independently evaluate the performance of automated total PINP assay and established age- and gender-specific reference intervals for PINP in healthy Korean population. METHODS: The imprecision, linearity, and detection capability of Elecsys total PINP assay was determined and reference interval was established using 599 serums from Korean population with normal bone mineral densities based on bone densitometry. Age groups were divided into 20s, 30s, 40s, 50s, 60s and over. RESULTS: Elecsys total PINP had excellent performance in imprecision, linearity, and detection capability. When partitioning age groups in Korean male and female populations, there was significant difference in total PINP between different age groups. In male populations, PINP level was decreased with increasing age, then it remained steady after middle-age. In female populations, there was a decreasing tendency similar to that in the male population with a sharp increase in the 50 to 59 age group. CONCLUSIONS: Elecsys total PINP assay showed precise and reliable performance in our study. We established age-related PINP reference intervals for Korean male and female population with normal bone mineral densities.

Measurement Uncertainty in Spine Bone Mineral Density by Dual Energy X-ray Absorptiometry

BACKGROUND: The purpose of this study was to calculate the measurement uncertainty of the process of bone mineral density (BMD) analysis using dual energy X-ray absorptiometry with traceability. METHODS: Between March 2015 and October 2016, among healthy participants in their 20s and 30s, the study included those who had not taken calcium, vitamin D supplements and steroids and were without a history of osteoporosis, osteopenia and diseases related to osteoporosis. Relational expression of the model was established based on Guide to the Expression of Uncertainty in Measurements and Eurachem and the uncertainty from each factor was evaluated. RESULTS: The combined standard uncertainty was 0.015, while the expanded uncertainty was 0.0298. The factor-specific standard uncertainties that occurred in the process of measuring BMD were 0.72% for the calibration curve, 0.9% for the internal quality control (IQC) using Aluminum Spine Phantom, 0.58% for European Spine Phantom (ESP), and 0.9% for the inspector precision (IP). CONCLUSIONS: The combined standard uncertainty of the spine BMD corrected with ESP was 0.015 when measured at one time and targeting one participant. The uncertainties of the accuracy of the IQC and the IP were higher than that of the other factors. Therefore, there will be a need for establishment of protocols to lower these uncertainties.

Clinical Usefulness of a DNA Microarray-based Assay for the Diagnosis of Sexually Transmitted Infections

BACKGROUND: Many molecular diagnostic methods have been developed to detect sexually transmitted infections (STI). The STDetect Chip (LabGenomics, Korea) which is a DNA microarray-based tool, newly developed for STI diagnosis in vitro, and the real-time PCR-based Anyplex STI-7 (Seegene, Korea) in clinical use were evaluated using ATCC DNA and clinical samples to determine the clinical usefulness of the STDetect Chip. METHODS: The two methods were compared for consistency, sensitivity, and specificity for 6 pathogens in 300 prospectively selected clinical samples. Analytical sensitivity for ATCC Neisseria gonorrhoeae, Chlamydia trachomatis, Mycoplasma hominis and Trichomonas vaginalis DNA and the effect of mixing bacterial DNA were studied. RESULTS: The consistency of the two methods for clinical samples was superior at more than 0.92 kappa value. The sensitivity and specificity of the STDetect Chip compared with Anyplex STI-7 were 90.5-98.8%, and 95.6-99.6%, respectively. With similar analytical performance for ATCC DNA, the STDetect Chip detected 10(-5) ng/µL of N. gonorrhoeae, 10(-4) ng/µL of C. trachomatis, 10(-6) ng/µL of M. hominis, and 10(-3) ng/µL of T. vaginalis. For the mixture of three bacterial DNAs, less sensitive detection level was observed for T. vaginalis. CONCLUSIONS: The STDetect Chip showed good agreement with the Anyplex STI-7 test and it is considered clinically useful for detecting sexually transmitted pathogens.

Result of Proficiency Test and Comparison of Accuracy Using a European Spine Phantom among the Three Bone Densitometries

BACKGROUND: Although dual energy X-ray absorptiometry (DXA) is known to standard equipment for bone mineral density (BMD) measurements. Different results of BMD measurement using a number of different types of devices are difficult to use clinical practice. The purpose of this study was to evaluate discrepancy and standardizations of DXA devices from three manufactures using a European Spine Phantom (ESP). METHODS: We calculated the accuracy and precision of 36 DXA devices from three manufacturers (10 Hologic, 16 Lunar, and 10 Osteosys) using a ESP (semi-anthropomorphic). The ESP was measured 5 times on each equipment without repositioning. Accuracy was assessed by comparing BMD (g/cm2) values measured on each device with the actual value of the phantom. Precision was assessed by the coefficient of variation (CVsd). RESULTS: Lunar devices were, on average, 22%, 8.3%, and 5% overestimation for low (L1) BMD values, medium (L2), and high (L3) BMD values. Hologic devices were, on average, 6% overestimation for L1 BMD, and 5% and 6.2% underestimation for L2 and L3 BMD values. Osteosys devices was, on average, 12.7% (0.063 g/cm2), 6.3% (0.062 g/cm2), and 5% (0.075 g/cm2) underestimation for L1, L2, and L3, respectively. The mean CVsd for L1-L3 BMD were 0.01%, 0.78%, and 2.46% for Lunar, Hologic, and Osteosys devices respectively. CONCLUSIONS: The BMD comparison in this study demonstrates that BMD result of three different devices are significant different between three devices. Differences of BMD between three devices are necessary to BMD standardization.

Discrepancy between Vitamin D Total Immunoassays due to Various Cross-reactivities

BACKGROUND: The purpose of this study was to find out the cause of discrepancy between various automated immunoassays for 25-hydroxy-vitamin D (25-[OH]D). METHODS: National Institute of Standards & Technology Standard Reference Material (SRM) 972a is SRM for 25-(OH)D and consists of 4 vials of frozen serum with different concentrations of 25-(OH)D. Each concentration was measured 6 times in 3 different immunoassays: ADVIA Vitamin D Total assay (Siemens Healthcare, Erlangen, Germany), ARCHITECT 25-(OH)D (Abbott Laboratories, Abbott Park, IL, USA), and COBAS Vitamin D Total assay (Roche Diagnostics, Basel, Switzerland). RESULTS: When using the certified reference values of SRM 972a as it is, discarding the cross-reactivity of each immunoassay, for ADVIA, the coefficient of determination (R2) as a score of regression analysis was 0.8995 and maximal difference between measured value and certified reference value was 3.6 ng/mL in level 3. The R2 and maximal differences of ARCHITECT were 0.5377 and 6.9 ng/mL, respectively, in level 4. Those of COBAS were 0.3674 and 22.3 ng/mL, respectively, in level 4. When considering cross-reactivities of each immunoassays to various 25-(OH)D metabolites, the ADVIA had R2 and maximal difference of 0.9254 and 3.3 ng/mL, respectively, in level 3. For ARCHITECT, the R2 and maximal differences were 0.7602 and 5.1 ng/mL, respectively, in level 1. Those of COBAS were 0.9284 and 4.9 ng/mL, respectively, in level 1. CONCLUSIONS: The cause of discrepancies between vitamin D immunoassays was mainly on the difference in cross-reactivities to various vitamin D metabolites. The discrepancies can be considerably decreased by considering cross-reactivities of each immunoassay.

Clinical Observation Study of Massive Blood Transfusion in a Tertiary Care Hospital in Korea

PURPOSE: Massive blood transfusios are uncommon. The goal of this study was to propose an ideal ratio for the blood component of massive hemorrhage treatment after review of five years of massive transfusion practice, in order to have the best possible clinical outcomes. MATERIALS AND METHODS: We defined a 'massive transfusion' as receiving 10 or more units of red blood cells in one day. A list of patients receiving a massive transfusion from 2004 to 2008 was generated using the electronic medical records. For each case, we calculated the ratio of blood components and examined its relationship to their survival. RESULTS: Three hundred thirty four patients underwent massive transfusion during the five years of the study. The overall seven-day hospital mortality for massive transfusion patients was 26.1%. Factors independently predictive of survival were a fresh-frozen plasma (FFP)/packed red blood cell (pRBC) ratio> or =1.1 with an odds ratio (OR) of 1.96 (1.03-3.70), and elective admission with an OR of 2.6 (1.52-4.40). The receiver operation characteristic (ROC) curve suggest that a 1 : 1 : 1 ratio of pRBCs to FFP to platelets is the best ratio for survival. CONCLUSION: Fixing blood-component ratios during active hemorrhage shows improved outcomes. Thus, the hospital blood bank and physician hypothesized that a fixed blood component ratio would help to reduce mortality and decrease utilization of the overall blood component.

HLA-DRB1 Study of DNA from Ancient Human Skeleton by Sequence-based Typing / 체질인류학회지

The analysis of ancient human DNA is increasingly used recently in the study of anthropology and human evolution. Although mitochondrial DNA and Y chromosomal DNA has commonly been the target in the field of human DNA study, HLA analysis of ancient human DNA is extremely rare. This study aimed to develop the PCR method of ancient human DNA for analyzing the sequence of HLA. Authors established a new method for HLA-DRB1 analysis by sequence-based typing. Alleles of HLA-DRB1 were analyzed and typed by sequencing with DNA of ancient human skeletons from Korea and Mongolia 3000-500 years ago. The types of HLA-DRB1 were determined by comparing the sequences with those of HLA database (http://www. ebi.ac.uk/Tools/blast2/nucleotide.html). The alleles of HLA-DRB1 of ancient human DNA from Korea and Mongolia were classified by types. The frequencies of HLA-DRB1 types of Mongolia were also presented according to the geography such as West, Central, East, and North. In summary, our method was successful in the analyzing the type of HLA-DRB1 from DNA of ancient human bones. Authors anticipate that many researchers could do their research in a better way to get the genetic information for the kinship analysis between individuals or communities from ancient human bones.

Annual Report on External Quality Assessment in Immunoserology in Korea (2009) / 임상검사와정도관리

The followings are the results for external quality assessment (EQA) in immunoserology for 2009: Evaluation of EQA was done in 2 trials in April and November, about 99% of laboratories participating average 7.4 items. The results were collected via internet and about 98% of laboratories have sent their results via internet. Control materials used in EQA were pooled sera including commercial controls, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA), which were delivered refrigerated for stability of control materials, being received within 48 hours after sending. Latex agglutination tests for rheumatoid factor (RF) showed frequently false positive or false negative results especially in commercial controls, possibly due to matrix effect. False negative and positive results were frequently found in the laboratories using immunochromatography assay (ICA) for anti-HCV and anti-HIV. More careful quality control should be required for ICA tests. New tests measuring non-treponemal and trponemal antibody such as turbidoimmunoassay (TIA) and chemiluminescence immunoassay (CLIA) were introduced. Standardization of instruments and reagents including calibrators for quantitative results should be required for the harmonization of results.

The Value of Procalcitonin and the SAPS II and APACHE III Scores in the Differentiation of Infectious and Non-infectious Fever in the ICU: A Prospective, Cohort Study

Early and accurate differentiation between infectious and non-infectious fever is vitally important in the intensive care unit (ICU). In the present study, patients admitted to the medical ICU were screened daily from August 2008 to February 2009. Within 24 hr after the development of fever (>38.3degrees C), serum was collected for the measurement of the procalcitonin (PCT) and high mobility group B 1 levels. Simplified Acute Physiology Score (SAPS) II and Acute Physiology And Chronic Health Evaluation (APACHE) III scores were also analyzed. Sixty-three patients developed fever among 448 consecutive patients (14.1%). Fever was caused by either infectious (84.1%) or non-infectious processes (15.9%). Patients with fever due to infectious causes showed higher values of serum PCT (7.8+/-10.2 vs 0.5+/-0.2 ng/mL, P=0.026), SAPS II (12.0+/-3.8 vs 7.6+/-2.7, P=0.006), and APACHE III (48+/-20 vs 28.7+/-13.3, P=0.039) than those with non-infectious fever. In receiver operating characteristic curve analysis, the area under the curve was 0.726 (95% CI; 0.587-0.865) for PCT, 0.759 (95% CI; 0.597-0.922) for SAPS II, and 0.715 (95% CI; 0.550-0.880) for APACHE III. Serum PCT, SAPS II, and APACHE III are useful in the differentiation between infectious and non-infectious fever in the ICU.

Genetic Association of Mitochondrial DNA Polymorphisms with Type 2 Diabetes Mellitus / 당뇨병

BACKGROUND: Although many single nucleotide polymorphisms (SNPs) of mtDNA have been found to be associated with type 2 diabetes mellitus, the results of studies using different population samples and different methods are mixed. Therefore, we conducted a genetic association study of mtDNA SNPs and type 2 diabetes mellitus in a Korean sample and compared our results with those of studies conducted in other human populations. METHODS: A total of 298 blood samples from 147 type 2 diabetic patients and 151 normal controls were surveyed for SNPs via PCR directed sequencing. Sequencing analyses were performed using the SeqMan module of the DNASTAR program. The identified SNPs were compared to previously reported SNP lists on NCBI and V-mitoSNP. RESULTS: A total of 24 SNPs were identified in the MT-RNR2, MR-TL1 and MT-ND1 mtDNA genes in Korean type 2 diabetes mellitus patients and normal controls. The SNPs identified in the Korean sample were not closely associated with the type 2 diabetes mellitus phenotype, a significantly different result from those previously observed in European, Chinese and Japanese samples. Additionally, a haplotype and prevalence analysis could not detect any differences between the type 2 diabetes mellitus patients and normal controls. CONCLUSION: The 24 mtDNA SNPs were not associated with type 2 diabetes mellitus risk in our Korean sample. The results of the present study support the possibility that mtDNA SNPs have a differential effect on the risk of type 2 diabetes mellitus according to geographical origin.

Annual Report on External Quality Assessment in Immunoserology in Korea (2008) / 임상검사와정도관리

The followings are the results for external quality assessment (EQA) in immunoserology for 2008:1.Evaluation of EQA was done in 2 trials in May and November, about 99% of laboratories participating average 7.7 items. The results were collected via internet and about 99% of laboratories have sent their results via internet. 2.Control materials used in the External Proficiecny Testing were pooled sera including Commercial controls, MAS(R) Immunology Control from Medical Analysis Systems (Camarillo, CA, USA), which were delivered refrigerated for stability of control materials, being received within 48 hours after sending. 3.Latex agglutination tests for rheumatoid factor (RF) showed frequently false positive or false negative results especially in Commercial controls, possibly due to matrix effect.4.False negative and positive results were frequently found in the laboratories using immunochromatography assay (ICA) for anti-HCV and anti-HIV. More careful quality control should be required for ICA tests. 5.New tests measuring non-treponemal and trponemal antibody such as turbidoimmunoassay (TIA) and chemiluminescence immunoassay (CLIA) were introduced.6.Standardization of instruments and reagents including calibrators for quantitative results should be required for the harmonization of results.

A Kinship Analysis of Ancient Human Bones and Teeth from Mongolia / 체질인류학회지

The kinship was analyzed genetically on the three 2000 year old ancient human bones and teeth excavated in Mongolia. The samples were processed in a clean room to prevent the contamination from modern human DNA. The DNA extraction and purification was done with ion-exchange column kit (Qiagen G-tip 20G, USA). The PCR was done with purified DNAs from ancient human bones for paternal Y-SNP haplogroup, maternal mtDNA haplogroup, and autosomal short tandem repeats (STR). Two samples belonged to the maternal D major haplogroup, which is one of the most frequent types in the present North East Asia. One of them, showing male genotype, belonged to the paternal C major haplogroup, which is also one of the most frequent types in the present North East Asia. The remaining one belonged to the paternal R major haplogroup, frequent in the present Europe, and the maternal U haplogroup, frequent in the present Europe and East Mediterranean. The repeated results were consistent in the autosomal STR PCR. The STR data were analyzed with DNA-VIEW program (http://www.dna-view.com), which showed no close kinship among the three ancient humans. Our method was successful in the analyzing kinship among ancient human bones, which has been possible in few restricted laboratories in the World. Authors anticipate that many researchers could do their research in a better way to get the genetic information from ancient human bones.

Metallo-beta-Lactamase-Producing Pseudomonas spp. in Korea: High Prevalence of Isolates with VIM-2 Type and Emergence of Isolates with IMP-1 Type

PURPOSE: Two Korean nationwide studies showed that metallo-beta-lactamases (MBLs)-producing-Pseudomonas spp. are not rare. The aim of this study was to assess the trends of MBL-producing isolates among imipenem-resistant isolates of Pseudomonas spp. MATERIALS AND METHODS: Imipenem-resistant clinical isolates were collected from 23 hospitals and one commercial laboratory participating in the KONSAR program in 2005. Polymerase chain reaction (PCR) was used to detect MBL genes. RESULTS: Alleles of MBL genes were detected in 10.8% of 415 Pseudomonas aeruginosa and 66.7% of 12 P. putida isolates from 18 of 24 hospitals/laboratory. Among the 14 IMP-1-like and 39 VIM-2-like MBLs, emergence of IMP-6 was detected for the first time. CONCLUSION: Prevalence of MBL-producing P. aeruginosa has not significantly increased, but IMP-6 emerged in P. aeruginosa.

Heteroplasmy of Mitochondrial DNA Shown in Korean and Mongolian / 체질인류학회지

As characterization of mitochondrial DNA (mtDNA) shows maternal inheritance and exists as more than thousands copies per cell, it is widely used for population genetics and forensic scientific field. However, mitochondrial DNA study has difficulties because heteroplasmy of mtDNA is being reported from coding and control region. In this study, we have analyzed 200 samples to examine heteroplasmy in mitochondrial DNA of Korean and Mongolian. The control region and coding region in mtDNA of blood from Koreans and Mongolians were analyzed with PCR amplication and sequencing. As a result, several heteroplasmy was observed from total 10 positions including 5 positions in coding region and 5 positions in control region, respectively. Moreover, it showed more than one heteroplasmy in coding region from 6 samples in Korean and 17 samples in Mongolian. Interestingly, heteroplasmy at 5178 position was shown in 6 samples among 23 samples. Considering that the position is important for deciding haplogroup D, we suggest that additional analysis on 4883 position needs for correct haplogrouping. Beside, we also found heteroplasmy in the other positions of 204, 4853, or 16249. Therefore, we suggest that it is required of combinatory analysis on several key nucleotide positions to obtain good results when determining mitochondrial haplogroups.

Comparison of SNP Occurred in Coding Regions of Mitochondrial DNA Analyzed from Korean and Mongolian / 체질인류학회지

Even though mitochondrial DNA analysis is performed in the field of molecular genetics, differences of the results exist regarding which nucleotide positions are analyzed. In this study, we strategically analyzed to find ethnic specific SNP of coding regions of mitochondrial DNA of Korean and Mongolian. Mitochondrial DNA was analyzed with PCR amplification and sequencing with 112 blood samples of Korean and 92 blood samples of Mongolian. As a result, the mutation which commonly appears both in Korean and Mongolian population is 17 nucleotide positions, and the one that shown in the only Korean is 13 nucleotide positions, the one that shown in the only Mongolian 26 nucleotide positions. However, it was thought as individual variation as most mutations are shown in a sample. Among them, it appears as 9% substitution rate in 10397, 4850 nucleotide position of Korean, whereas 12.3% or 15% substitution rate in 5108, 9950 nucleotide positions of Mongolian, respectively. Beside, we observed high level of heteroplasmy in 3546, 3553 nucleotide positions. Therefore, we suggest that these regions might be novel genetic markers for dividing mitochondrial haplogroup of Korean and Mongolian population, but additional analysis needs on several nucleotide positions in huge samples as analyzing on restricted nucleotide positions using restricted DNA samples.

Annual Report on External Quality Assessment in Immunoserology in Korea (2007) / 임상검사와정도관리

The followings are the results for external quality assessment (EQA) in immunoserology for 2007: 1. Evaluation of EQA was done in 2 trials in May and December, about 99% of laboratories participating average 7.8 items. The results were collected via internet for the first time and 96~98% of laboratories have sent their results via internet. 2. All the specimens for Immunoserology in EQA were delivered refrigerated, being received within 48 hours after sending. 3. Commercial controls, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA) were used to assure the quality of quantitative results of C-reactive protein (CRP), rheumatoid factor (RF) and anti- streptolysin O (ASO) tests, and the RF results of MASR Immunology Control were variable depending on the reagents used. 4. The laboratories using immunochromatography assay (ICA) were increased, however, many laboratories using ICA reported falsely negative for the positive specimens. The sensitivity of ICA test kits as well as various factors influencing the ICA results should be evaluated. 5. The HBsAg results of the ACCURUN 1R Multi-Marker Positive Control (Boston Biomedica Inc. USA) were falsely reported as negative in some laboratories using arbitrarily determined cutoff. 6. Standardization of methods including calibrators for quantitative results should be required for the harmonization of results.

Annual Report on External Quality Assessment in Immunoserology in Korea (2007) / 임상검사와정도관리

The followings are the results for external quality assessment (EQA) in immunoserology for 2007: 1. Evaluation of EQA was done in 2 trials in May and December, about 99% of laboratories participating average 7.8 items. The results were collected via internet for the first time and 96~98% of laboratories have sent their results via internet. 2. All the specimens for Immunoserology in EQA were delivered refrigerated, being received within 48 hours after sending. 3. Commercial controls, MASR Immunology Control from Medical Analysis Systems (Camarillo, CA, USA) were used to assure the quality of quantitative results of C-reactive protein (CRP), rheumatoid factor (RF) and anti- streptolysin O (ASO) tests, and the RF results of MASR Immunology Control were variable depending on the reagents used. 4. The laboratories using immunochromatography assay (ICA) were increased, however, many laboratories using ICA reported falsely negative for the positive specimens. The sensitivity of ICA test kits as well as various factors influencing the ICA results should be evaluated. 5. The HBsAg results of the ACCURUN 1R Multi-Marker Positive Control (Boston Biomedica Inc. USA) were falsely reported as negative in some laboratories using arbitrarily determined cutoff. 6. Standardization of methods including calibrators for quantitative results should be required for the harmonization of results.

Sex of Ancient Mongolian Human Bones Using Biallelic Marker RPS4Y for Y haplogroup / 대한해부학회지

Many data from ancient human remains became useful by molecular approach for ancient human DNA. In anthropology, genetic sex is essential to understand marriage and burial patterns, differential mortality rates between sexes, and differential patterns by sex of disease, diet, status, and material possessions. This study was designed to determine genotype sex of 52 ancient human bones with well preserved skulls, and to compare with the orphological sex. Parts of femur and other bones were used as ancient bones excavated in Mongolia aged between bronze and Mongol period. Morphological sex was determined by Mongolian scientist, and genotype sex was determined by using biallelic marker RPS4Y for Y haplogroup. Of 52 genetic males, 10 samples were morphologically female. In conclusion, biallelic marker RPS4Y. PCR amplication method will be useful in sex determination of ancient bones.

Comparison of an Immature Platelet Fraction and Reticulated Platelet in Liver Cirrhosis / 대한진단검사의학회지

BACKGROUND: The main causes of thrombocytopenia in cirrhosis are thought to be platelet destruction and the reduction of thrombopoietin (TPO) expression in the liver. Immature platelet fraction (IPF) has been measured by a fully automated analyzer (Sysmex XE-2100, Japan) as reticulated platelet (RP), which is reflected with thrombopoiesis in bone marrow. In this study, we tried to compare the percentage of IPF (IPF) with that of RP (RP) in patients with liver cirrhosis (LC) and controls. METHODS: We compared IPF to RP in 72 liver cirrhosis patients and 30 healthy normal controls. RP was stained with acridine orange, followed by FC500 (Beckman Coulter, USA) analysis and the IPF was identified by flow cytometry with the use of a nucleic acid specific dye in the reticulocyte channel on the Sysmex XE-2100 (TOA Medical Electronics Co., Ltd., Japan). RESULTS: IPF value in the healthy control was 2.2% (1.7-5.2). RP and IPF were significantly higher in the patients with liver cirrhosis (P<0.05). IPF appeared to be correlated with RP (y=0.19x+3.35, r=0.34, P<0.05). In ROC for diagnosis of LC, IPF was significantly more useful than RP. CONCLUSIONS: This results show that a rapid, inexpensive automated method for measuring the IPF is feasible and should become a standard parameter in evaluating reticulated platelets.

Comparison between Morphological Sex and Genotype Sex of Uzbekistan Ancient Bones Using Improved Amelogenin PCR Amplication Method / 체질인류학회지

Determination of male and female is important in anthropology, archeology and forensic science. This study was designed to compare genotype sex of improved amelogenin PCR amplication method with morphological sex of ancient human bones. Sixty human skulls which lived from the Bronze Age to twenties centuries and excavated in Uzbekistan were used in this study. Morphological sex was determined by Uzbekistan scientist, and genotype sex was determined by improved amelogenin PCR amplication developed in this study. Among 20 morphological males, 13 samples (65%) were genotypical male. Among 40 morphological females, 20 samples (50%) were genotypical male. In conclusion, morphological method might be inadequate for sex determination of ancient bones. The improved amelogenin PCR method will be useful in sex determination of ancient bones.