Simple Identification Method Research Articles

Multiplexed Analysis of Photochemical Oxidants Using a Nanoparticle-Based Optoelectronic Nose.

Photochemical pollutants pose a substantial threat to human health in both outdoor and indoor environments. Herein, we prepare a class of gold nanoparticle-based colorimetric sensor arrays on optimized hydrophobic substrates using a simple pin-printing method for accurate identification and quantification of various gas-phase oxidants, as these microdetectors are low cost, sensitive, and easy to fabricate. For an array of AuNP sensors modified with various thiol-terminated ligands, a unique and distinguishable change in color (i.e., red, green, and blue response patterns) was obtained for each specific pollutant for molecular fingerprinting. Remarkable discrimination among 15 gases at a fairly low vapor concentration (i.e., 500 ppb) was illustrated using standard chemometric methods. Using digital imaging, the AuNP colorimetric sensor array offers ultrasensitive dosimetric identification of gas-phase oxidants relevant to outdoor and indoor air pollution, with limits of detection generally at sub-ppb levels for 2 h measurement. As a practical application, the sensor array is able to predict the overall air quality in indoor office environments over 24 h. Such sensor array based on chemically induced sintering of nanoparticles has significant implications for the development of nanosensors used in continuous monitoring of potential airborne pollutants at low concentrations from a large number of locations in a cost-effective manner.

A simple and efficient method for isomorphism identification of planar kinematic chains

Isomorphism identification plays an important role in structural design and innovative design. Based on the adjacency matrix and loop theory, a new method is proposed in this paper to identify the isomorphic kinematic chains. It enriches the application of loop-based theory for isomorphism identification. In the kinematic chain, links and joints are connected alternatively and every link corresponds to a fixed link degree. Due to the inherent characteristics, the labeled sequence of links can be random, which does not affect the result of isomorphism identification. By the programming software MATLAB, some examples with 6-, 8-, 10-, 11-, 12-link kinematic chains, and 15-vertex topological graphs are presented. Results show that the proposed method applies to topology graphs and kinematic chains with one or multiple joints. Compared with other methods, the proposed method is confirmed correctly. And there is no counterexample. It lays a solid foundation for structural synthesis in the future.

A Simple Method of Intraoperative Identification of a Reliable Perforator while Dissecting an Anterolateral Thigh Flap.

A Simple Method of Intraoperative Identification of a Reliable Perforator while Dissecting an Anterolateral Thigh Flap.

Development of an inoculation technique for rapidly evaluating maize inbred lines for resistance to stalk rot caused by Fusarium spp. in the field.

Maize (Zea mays L.) stalk rot, caused primarily by the soil-borne fungal pathogen Fusarium spp., reduces maize quality and yield worldwide. This study was undertaken to develop and utilize a rapid continuous injection inoculation technique to evaluate maize inbred lines for resistance to Fusarium spp. under field conditions, which could facilitate the identification and development of new sources of host resistance to manage the disease. Continuous injection inoculation is a rapid, stable, and simple method that can evaluate the resistance of maize inbred lines to Fusarium stalk rot (FSR) within 20 days. To verify the feasibility and reliability of inoculation method, Fusarium graminearum, F. proliferatum, and F. subglutinans were isolated, identified, and inoculated into maize at the six-leaf stage (V6) by a veterinary adjustable bottle continuous vaccination syringe. Our results showed that out of a total of 97 inbred maize lines, six (6.2%) showed high resistance to maize stalk rot, 20 showed resistance (20.6%), 32 were susceptible (33.0%), and 39 were very susceptible (40.2%). Based on simple sequence repeat (SSR) markers, an analysis of molecular variance indicated a significant correlation between population of the inbred maize line and resistance to FSR (P = 0.001). Overall, this study provided a systematic, rapid, stable, and simple identification method for maize inbred lines resistant to FSR in the field. At the same time, this method was also suitable for genetic diversity analysis of maize inbred lines resistant to FSR.

A rapid LAMP‐based colorimetric assay with quick DNA extraction for on‐site identification of Drosophila suzukii Matsumura

AbstractThe spotted wing drosophila, Drosophila suzukii Matsumura (Diptera: Drosophilidae), distributes in most soft‐skinned fruit areas in China, is an economically important pest of fresh cherries during Sino‐European trade and is considered a quarantine pest in A2 list by European and Mediterranean Plant Protection Organization. Loop‐mediated isothermal amplification (LAMP) is an accurate, quick and convenient molecular identification method, applied to distinguish D. suzukii from other fruit flies. This method can be used for inspection in the field, and at the points of entry (POEs), because its results can be detected with the naked eye due to colour changes. In our study, first, we reported a simple and fast LAMP colorimetric detection method for molecular identification of D. suzukii. We designed primer sets for D. suzukii based on mitochondrial cytochrome oxidase I (COI) sequences. The specificity and sensitivity of the primers were tested by using target and non‐target fruit flies in the family Drosophilidae and Tephritidae, usually intercepted during the Sino‐European cherry trade. Second, for detection in the field and at POEs, the adoption of a quick DNA extraction method could save the total time of LAMP identification to about 90 min. Taken together, this accurate, quick and convenient LAMP‐based colorimetric identification assay combined with a quick DNA extraction method could visually detect clearly with just one portable heating device, which will be useful for rapid on‐site identification and inspection for D. suzukii by the quarantine department.

NMR Study of the Discrimination of Enantiomers of Methamphetamine and Its Raw Materials Using Chiral Solvating Agents

Some controlled substances, such as stimulants and narcotics, have asymmetric carbons in their molecules. Because the enantiomers do not always show the same pharmacological effects, and there are substances with different controls due to differences in their stereochemistry, a simple and unambiguous method for assessment of the composition of enantiomers is necessary. In this study, to develop a simple and rapid stereoscopic identification method for methamphetamine and its raw materials (ephedrine and pseudoephedrine), the 1H-NMR method was studied using three commercially available chiral solvating agents (CSAs); 1,1'-bi(2-naphthol)(BINOL), 2,2,2-trifluoro-1-(9-anthryl)ethanol (TFAE) and α-methoxy-α-(trifluoromethyl)phenylacetic acid (MTPA). In addition, the accuracy of the optical purity, which was measured using samples mixed with enantiomers in various ratios, was investigated. The NMR peaks of the enantiomers were separated by adding (R)- or (S)-form of BINOL, TFAE or MTPA to the chloroform-d solution of methamphetamine, ephedrine or pseudoephedrine. A sufficient discrimination of enantiomers was obtained by adding about 10 equal amounts of each CSA to the solutions. With regard to the optical purity, it was possible to determine accurately the mixing of small amounts of enantiomers of about 5% even if the NMR peaks did not reach the baseline separation, when impurity peaks do not overlap. This method will be one of the useful techniques for the rapid and simple discrimination of enantiomers of illegal methamphetamine and its raw materials.

Identification of Early-Stage Cervical Cancer Tissue Using Metamaterial Terahertz Biosensor With Two Resonant Absorption Frequencies

We propose a simple method for fast and label-free identification of early-stage cervical cancerous tissues using a metamaterial (MM) terahertz (THz) biosensor with two resonant absorption frequencies at 0.286 THz and 0.850 THz, respectively. Because the two resonant absorption frequencies are very sensitive to the ambient change of dielectric properties, the cervical cancerous tissues can be distinguished from the normal tissues by analyzing the resonant frequency shift. The calculated sensitivity of the low resonant frequency and that of the high resonant frequency are 29 and 74 GHz/RIU, respectively. In the experiment, the cervical cancerous tissue samples were taken from different patients, and the cancerous and normal parts were determined by hematoxylin-eosin (HE) stain. Compared with the traditional method of directly detecting cancer tissue without the sensor, this method has the advantages of being simple, label free and fast. In addition, compared with biosensors which have a single resonant absorption frequency, the biosensors with double resonant absorption frequencies have higher accuracy. This method potentially can be applied for the detection and diagnosis of many other cancer tissues.

Validation of salivary ferning based estrus identification method in a large population of water buffaloes (Bubalus bubalis) using Foldscope

Missing an estrus event is an economic problem in buffaloes because of lack of a simple and accurate estrus identification method. Saliva, a non-invasive fluid available every time, showed typical fern-like crystallization patterns at early estrus in buffaloes. However, to implement this salivary ferning based estrus identification method in the field conditions, the present study, for the first time, validated this method in four buffalo population samples (PS) representing four different field scenarios, an organized herd (PS1, 10 buffaloes monitored daily for a year (149 estrus events)), artificial insemination (AI) centers (PS2, 114 buffaloes brought for AI), induced estrus condition (PS3, 44 buffaloes) and farmers’ doorsteps (PS4, 275 random buffaloes with unknown reproductive history and no estrous signs). Totally, 582 saliva samples were collected from 443 buffaloes. Salivary smears were observed under a simple microscope and/or a less expensive (< 1USD) paper microscope, Foldscope. On the basis of salivary fern-like patterns, the proportions of estrus identification were significantly different among PS. Specifically, the proportions in the PS1 (0.74, P < 0.0001) and PS4 (0.08, P < 0.05) were significantly higher than their population proportion estimates, 0.5 and zero, respectively. Therefore, this estrus identification method is much useful in the PS1 and PS4, the practical field scenarios requiring an accurate estrus prediction method, compared to the PS2 and PS3. Especially, this method is 91 % confirmatory to detect early estrus in PS4. Therefore, salivary ferning is a useful technique to identify early estrus in buffaloes in the field conditions at farmers’ doorsteps.

A novel strategy of "pick the best of the best" for the nondestructive identification of Poria cocos based on near-infrared spectroscopy.

In this paper, a novel strategy of “pick the best of the best” was proposed for the nondestructive identification of different‐origin and adulterated Poria cocos with near‐infrared spectroscopy. First, various preprocessing methods were divided into three classes: baseline correction, scattering and trend correction, and scaling. The single preprocessing methods with the best predictions in each class were selected. Then, the selected preprocessing methods were combined in pairs according to three classes. The pair combination preprocessing methods with the best predictions and also better predictions than single methods were selected. Finally, the selected pair combination preprocessing method was combined with the methods in the unselected class. The three combination preprocessing methods with the best predictions and also better predictions than pair combination methods were selected as the final prediction. With this strategy, the optimized preprocessing combination can be obtained quickly, and the identification accuracy with principal component analysis method can be greatly improved. 0% identification accuracy of adulterated samples and 12.5% identification accuracy of different‐origin samples were obtained with the raw data. However, 100% accuracy of adulterated samples, 93.8% accuracy of calibration dataset, and 75% accuracy of validation dataset can be obtained with the novel strategy. The developed technology can be regarded as a simple, rapid, and accurate nondestructive identification method for different‐origin and adulterated samples, and has a broad application prospect in the future.

Development and optimization of a method based on QuEChERS-dSPE followed by UPLC-MS/MS for the simultaneous determination of 21 mycotoxins in nutmeg and related products

Nutmeg (Myristica fragrans), a popular spice and traditional medicine worldwide, is susceptible to fungal and mycotoxin contamination. Hence, it is important to monitor the residual levels of multiple mycotoxins in nutmeg and related products. However, analysts are continuously faced with the challenges of quantifying mycotoxin concentrations in complex matrices such as spices and traditional Chinese medicines. To develop a simple and sensitive mycotoxin analysis method encompassing the total workflow from sample preparation to quantitative analysis, a simple, robust, and sensitive method for the simultaneous identification and quantification of 21 mycotoxins in nutmeg and related products was described. The method is based on sample preparation by modified QuEChERS extraction and dispersive solid-phase extraction (dSPE) cleanup followed by ultrahigh-performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-MS/MS). Matrix-matched calibration was applied for the quantification of the investigated mycotoxins. The limits of detection of the target analytes ranged from 0.04 μg/kg to 6.0 μg/kg. Recovery rates ranged from 60.49% to 97.46% at three spiking levels, with relative standard deviations<13%. This method was applied to 45 samples of nutmegs, mace and related products. Three nutmeg samples and one nutmeg powder were contaminated with aflatoxins with residue levels of 1.76 ~ 3.39 μg/kg for AFB1 and 2.51 ~ 8.77 μg/kg for AFs (the sum of AFB1, AFB2, AFG1, AFG2 and AFM1). No mycotoxins were detected in mace. All of the contamination levels were below the regulatory maximum residue limits suggested by the European Commission and China.

Rapid detection of avian leukosis virus subgroup J by cross-priming amplification

Avian leukosis virus subgroup J (ALV-J) causes oncogenic disease in chickens in China, resulting in great harm to poultry production, and remains widespread in China. Herein, we employed a cross-priming amplification (CPA) approach and a nucleic acid detection device to establish a visual rapid detection method for ALV-J. The sensitivity of CPA, polymerase chain reaction (PCR) and real-time PCR (RT-PCR) was compared, and the three methods were used to detect ALV-J in the cell cultures which inoculated with clinical plasma. The result showed when the amplification reaction was carried out at 60 °C for just 60 min, the sensitivity of CPA was 10 times higher than conventional PCR, with high specificity, which was comparable with RT-PCR, based on detection of 123 cell cultures which inoculated with clinical plasma, the coincidence rate with real-time PCR was 97.3% (71/73). CPA detection of ALV-J does not require an expensive PCR instrument; a simple water bath or incubator is sufficient for complete DNA amplification, and the closed nucleic acid detection device avoids aerosol pollution, making judgment of results more intuitive and objective. The CPA assay would be a promising simple, rapid and sensitive method for identification of ALV-J.

Development and Validation of a Effective and Reliable HPLC Method for the Quantification of Levodopa and Carbidopa in Pharmaceutical Formulations

A convenient and simple high performance liquid chromatography method for simultaneous separation, determination and identification of levodopa and carbidopa in pharmaceutical formulation has been developed. The analysis was carried out using Ace C18 (4.6 x 250 mm, 5 µm) column, and the separation was performed using a mobile phase consisting of 50 mM KH2PO4 (pH 2.3) pumped at a flow rate of 1.2 mL min-1 with UV detection at 280 nm. The method has been successfully validated on the basis of the International Conference on Harmonization (ICH) acceptance criteria. The method is selective, since no interferences corresponding to these analytes were found at retention times. Retention times for both drugs were 2.939 min and 5.018 for levodopa and carbidopa, respectively. The method was validated and shown to be linear in the concentration range of 25-125 µg mL-1 and 6.25-31.25 µg mL-1 for levodopa and carbidopa, respectively. The method exhibited good linearity (R2 &amp;gt; 0.999) over the assayed concentration range and showed good intra-day and inter-day precision. The limit of detection (LOD) and limit of quantification (LOQ) were determined based on standard deviation of the intercept and the slope of the calibration curve. LOD and LOQ values were 0.70 µg mL-1 and 2.13 µg mL-1 for levodopa and 1.31 µg mL-1 and 3.96 µg mL-1 for carbidopa, respectively. The method's accuracy was tested by adding known volume of standard solution (75 percent, 100 percent, and 125 percent concentration of the sample) to the 100 percent concentration pre-analyzed sample solution. The percentage mean recovery by standard addition experiments of levodopa and carbidopa is 99.75% and 99.55%, respectively. This method has also been successfully used for the determination of levodopa and carbidopa in pharmaceutical formulations.

A simple and rapid method for fish sex identification based on recombinase-aided amplification and its use in Cynoglossus semilaevis

Fish sex identification is a basic technique of great importance for both fish genetic studies and fisheries. Due to the sexual reversal phenomenon in many fish species, a simple and rapid molecular identification method for fish genetic sex is urgently needed to suit versatile detection scenarios, such as point-of-need applications. In this study, we took Cynoglossus semilaevis as an example, established a recombinase-aided amplification (RAA)-based method for sex identification, and combined the RAA-detection with two result visualization approaches with distinct features, capillary electrophoresis (CE) and lateral flow dipstick (LFD). Specific primers and probe were designed to specifically detect the sex chromosome W of C. semilaevis in order to distinguish the genetic sex between males, pseudo-males and females. To evaluate the performance of our methods, the genetic sex for twenty-eight males, sixty-eight pseudo-males and fifty-four females were examined with the RAA-based method and classical PCR-based genotyping method, demonstrating the consistent results of sex identification between both methods. The RAA-LFD method is operationally simple, rapid (~ 30 min) and holds great potential for point-of-need applications of fish sex identification, including fishery fields. The method presented here could be effective for identifying fish gender with the ZW karyotype.

Educational tool for analysis of proportional integral and fractional order proportional integral controlled quadratic boost converter system using MATLAB/simulink

This paper displays an simple method for identification of controller for wind based Quadratic Boost Converter Inverter system. India, being the fourth largest wind power generator has attracted numerous researchers towards the improvement of wind energy conversion system. This paper also presents improved controller techniques for a Permanent Magnet Synchronous Generator (PMSG) coupled with Cascaded Quadratic Boost Converter (QBC) and Space Vector Modulation Inverter (SVMI). The digital simulation and execution of PMSG based wind turbine along with QBC and SVM Inverter is presented in a closed loop system. The performance of closed loop system is realized using Proportional Integral (PI) and Fractional Order Proportional Integral (FOPI) controllers. Initially, the AC power from PMSG wind turbine is rectified to DC using rectifier circuit. The DC power from the bridge rectifier is then boosted to the required level using quadratic boost converter. The output from QBC is then given to the SVM inverter. The closed loop investigations are carried with PI and FOPI controllers. The simulation results of both PI and FOPI controlled QBC are compared. The outcome of FOPI controller represents that the steady state error and settling time are reduced when compared to PI controlled closed loop Quadratic Boost Converter. The overall Matlab/simulink model is applied to undergraduate/postgraduate course as a educational tool and assessed thoroughly.

A novel non-invasive identification of genome editing mutants from insect exuviae.

With the wide application of genome editing in insects, a simple and efficient identification method is urgently needed to meet the increasing demand for mutation detection. Here, taking migratory locusts as a model system, we developed a non-invasive method to accurately identify genome-edited mutants by using DNA from insect exuviae. We compared the quantity and quality of genomic DNA from exuviae in five instar hoppers and found that the 1st instar exuviae had the highest DNA yield and content, while the 3rd instar exuviae had the best quality. Consensus genotypes were identified from genomic DNA of hoppers at different developmental stages in the same individuals. Moreover, we demonstrated that the amplification products from DNA extracted from locust exuviae are the consensus sequences with those from the hemolymph and foreleg pre-tarsus. Therefore, non-invasive samples provide the same genotyping results as minimally invasive and invasive samples of the same individuals. Furthermore, this identification method that uses genomic DNA from exuviae can be used for early screening of positive genome-edited individuals in each generation for adult crossing. In our study, the non-invasive identification method was not only simpler and provided results earlier than existing methods, but also had a better reproducibility and accuracy. This non-invasive identification approach using genomic DNA from exuviae can be adapted to meet the growing demand for genetic analysis and will find wide application in insect genome editing research.

Wing Morphometrics of Aedes Mosquitoes from North-Eastern France.

Simple SummaryMosquitoes act as vectors of arboviruses and their correct identification is very important to understanding the diseases they transmit. To date, this identification is based on several techniques that are either expensive or time consuming. Wing geometric morphometrics allow fast and accurate mosquito identification. By analyzing the pattern of wing venation, it is possible to separate mosquito species. We applied this technique on six Aedes mosquito species from north-eastern France. Our results show a very good differentiation of these species. The use of wing geometric morphometrics could increase the efficiency of field entomologists in case of viral outbreaks. Integrated with existing morphological identification software, it might help relocate mosquito identification from the lab to the field.Background: In the context of the increasing circulation of arboviruses, a simple, fast and reliable identification method for mosquitoes is needed. Geometric morphometrics have proven useful for mosquito classification and have been used around the world on known vectors such as Aedes albopictus. Morphometrics applied on French indigenous mosquitoes would prove useful in the case of autochthonous outbreaks of arboviral diseases. Methods: We applied geometric morphometric analysis on six indigenous and invasive species of the Aedes genus in order to evaluate its efficiency for mosquito classification. Results: Six species of Aedes mosquitoes (Ae. albopictus, Ae. cantans, Ae. cinereus, Ae. sticticus, Ae. japonicus and Ae. rusticus) were successfully differentiated with Canonical Variate Analysis of the Procrustes dataset of superimposed coordinates of 18 wing landmarks. Conclusions: Geometric morphometrics are effective tools for the rapid, inexpensive and reliable classification of at least six species of the Aedes genus in France.

Penggunaan KOH String Test Sebagai Alternatif Identifikasi Awal Bakteri Gram Negatif

KOH string test is a simple identification method without staining, requiring only one reagent to determine the Gram reaction in bacteria. Considering the simplicity of this method, its use needs to be introduced further. However, to ensure the validity of this method, research needs to be carried out to verify the results of the KOH string test when compared with the identification method using Gram staining, which is the gold standard. This study aimed to determine whether there is a difference in the identification results between Gram staining and the KOH string test in determining the Gram characteristics of bacteria, especially Gram-negative bacteria, to find out whether there is a difference in the length of mucus produced from the KOH string-test between KOH reagent concentrations of 1, 3, and 5%, as well as knowing the KOH concentration that was best used for bacterial identification with the KOH string test. The results of the study showed that there was no difference in results between identification using Gram staining and the KOH string test. Statistical results showed a significant difference between the mucus lengths of the three reagent concentrations, and 3% KOH was the recommended reagent for use in the KOH string test method.

Genetic Identification of Edible Bird's Nest in Thailand Based on ARMS-PCR.

Edible bird’s nest (EBN) is a popular delicacy in the Asian Pacific region originating from Indonesia, Malaysia, Thailand and Vietnam, which consist of various potential medicine value in Traditional Chinese Medicine (TCM). Thailand is one of the main exporters of EBN. However, the genetic information of EBN, a key part of molecular biology, has yet to be reported in Thailand. It is necessary to explore the genetic information of EBN in Thailand based on a quick and simple method to help protect the rights and interests of consumers. This research aimed to systematically evaluate different methods of extracting EBN DNA to improve the efficiency of the analysis of cytochrome b (Cytb) and NADH dehydrogenase subunit 2 (ND2) gene sequences, the establishment of phylogenetic trees, and the genetic information of EBN in Thailand. Additionally, we aimed to develop a quick and simple method for identifying EBN from different species based on the genetic information and amplification-refractory mutation system PCR (ARMS-PCR). By comparing the four methods [cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), kit and guanidinium isothiocyanate methods] for EBN extraction, we found that the guanidinium isothiocyanate method was the optimal extraction method. Phylogenetic trees generated on the basis of Cytb and ND2 gene analyses showed that 26 samples of house EBN and 4 samples of cave EBN came from Aerodramus fuciphagus and Aerodramus maximus, respectively. In addition, to distinguish different samples from different species of Apodiformes, we designed 4 polymerase chain reaction (PCR) amplification primers based on the ND2 gene sequences of A. fuciphagus and A. maximus. The ARMS-PCR results showed band lengths for A. fuciphagus EBN of 533, 402, and 201 bp, while those for A. maximus EBN were 463, 317, and 201 bp. Collectively, the results showed that ARMS-PCR is a fast and simple method for the genetic identification of EBN based on designing specific original identification primers.

Simplified rapid differentiation of Liriomyza chinensis Kato (Diptera; Agromyzidae) genotypes A and B using PCR‐RFLP

AbstractThe stone leek leafminer Liriomyza chinensis Kato is one of the most important pests of Allium spp. In recent years, a new genotype of L. chinensis, genotype B, has been shown to cause more severe damage to Allium spp. compared with the native genotype A in Japan. Thus, identification of the genotypes is important for establishing L. chinensis control strategies. In this study, we developed a simple and rapid method of genotype identification using PCR‐restriction fragment length polymorphism (RFLP). Primers were designed to mitochondria cytochrome oxidase subunit I gene (mtCOI) nucleotide sequences from both genotypes. Genotypes A and B were successfully identified using the restriction enzymes PvuI and DdeI, respectively. Using PCR‐RFLP, we identified the genotype of L. chinensis samples (n = 108) collected in seven Japanese bunching onion fields in Ibaraki Prefecture, Japan. All individuals were shown to be genotype B, as expected.

A simple PCR-based genetic sex identification method in the blotched snakehead (Channa maculata) developed by high-throughput sequencing

Sex dimorphism is a common phenomenon in animals. Blotched snakehead (Channa maculata) is an economically important freshwater fish with a faster growth rate in males. However, existing sex-specific markers were too few and difficult to use in production practice, which hindered the development of all-male breeding. Here, we generated 15.64, 20.77 and 29.03 Gb paired-end genomic sequence reads for three male libraries, 12.82, 19.36 and 28.33 Gb for three female libraries and 30.93 Gb for a female mixed pool libraries to screen male-specific sequences. After assembly, we obtained a male reference genome with 541,932 assembled sequences and a female reference genome containing 853,706 assembled sequences. After mapping, 23 male-specific sequences were identified. Six candidate male-specific markers were successfully validated using PCR reaction. Moreover, based on homology analysis and coverage information, we found a candidate X chromosome sequence that was partially similar to the corresponding Y chromosome sequence, and two resulting primers were successfully used in the identification of female (XX), male (XY) and supermale (YY) individuals. In general, a deep genome sequencing method was first provided to effectively enrich large sex-specific segments of Blotched snakehead, which would greatly contribute to elucidation of its sex determination mechanisms and development of all-male breeding.